Cyclic ether derivatives of pyrazolo[1,5-a]pyrimidine-3-carboxyamide

ABSTRACT

The invention relates to Spirocyclic ether derivatives of pyrazolo[1,5-a]pyrimidine-3-carboxyamide of formula (I) 
     
       
         
         
             
             
         
       
     
     wherein R 1 , R 2 , R 3 , R 5 , R 6  and A are as defined herein. The compounds of formula (1) are inhibitors of phosphodiesterase 2 and useful in treating central nervous system diseases and other diseases. The invention relates to processes for preparing pharmaceutical compositions as well as processes for manufacture the compounds according to the invention.

FIELD OF THE INVENTION

The invention relates to cyclic ether derivatives ofpyrazolo[1,5-a]pyrimidine-3-carboxyamide of general formula (I) whichare inhibitors of phosphodiesterase 2 and useful in treating centralnervous system diseases and other diseases.

In addition, the invention relates to processes for preparingpharmaceutical compositions as well as processes for manufacture thecompounds according to the invention.

BACKGROUND OF THE INVENTION

Phosphodiesterase 2 (PDE2) inhibitors are promising therapeutic targetsfor treatment of cognitive impairment in diseases such as Schizophrenia,Alzheimer's disease and depression. Inhibitors of PDE2 have emerged aspotential candidates to improve synaptic plasticity and memory function.

Phosphodiesterases (PDE) are expressed in nearly all mammalian cells. Todate eleven families of phosphodiesterases have been identified inmammals. It is well established that PDEs are critically involved incell signalling. Specifically, PDEs are known to inactivate the cyclicnucleotides cAMP and/or cGMP.

PDE2 hydrolyses both, cGMP and cAMP. It is both abundantly expressed inthe brain indicating their relevance in CNS function.

The expression of PDE2 in the hippocampus, the cortex and in thestriatum indicate an involvement in the mechanism of learning andmemory/cognition. This is further supported by the fact that increasedlevels of both cGMP and cAMP are involved in the process of short andlong term potentiation (LTP) forming. Further data support theprocognitive effect of PDE2 and a synergistic effect of PDE2 oncognition.

Furthermore, the expression of PDE2 in the nucleus accumbens (part ofthe striatum), the olfactory bulb, the olfactory tubercle and theamygdala supports additional involvement of PDE2 in the pathophysiologyof anxiety and depression. This is supported by in vivo studies.

It is commonly accepted (free drug hypothesis) that unbound or free drugconcentration at the site of action is responsible for pharmacologicalactivity in vivo at steady state and, in the absence of activetransport, the free drug concentration is the same in any biomembrane.

For drugs with an intended action in the central nervous system (CNS),it is assumed that unbound drug in interstitial spaces (ISF) in thebrain is in direct contact or in equilibrium with the site of action.Because cerebrospinal fluid (CSF) is in direct contact with the braintissue, it is assumed to readily equilibrate with brain interstitialfluid concentration so that CSF concentration is used as a commonsurrogate measure for drug unbound concentration in pre-clinicalpharmacology studies. Accordingly, for compounds with an intended actionin the central nervous system it is important that they reach a high CSFconcentration and a high CSF to plasma ratio in order to have highpharmacological activity in the CNS.

At steady state and in the absence of active transport, the unboundbrain concentration can also be estimated with the experimentally moreaccessible unbound plasma concentration by measuring the plasma proteinbinding (PPB) across species.

High membrane permeability and absence of active transport process atthe BBB (blood brain barrier) together with plasma/brain tissue bindingare recognised as the primary determinant of drug disposition withinCNS.

High metabolic stability is desirable in order to achieve significantexposure of a drug within the body.

Several families of PDE2 inhibitors are known. Imidazotriazinones areclaimed in WO 2002/068423 for the treatment of e.g. memory deficiency,cognitive disorders, dementia and Alzheimer's disease. Oxindoles aredescribed in WO 2005/041957 for the treatment of dementia. Furtherinhibitors of PDE2 are known from WO 2007/121319 for the treatment ofanxiety and depression, from WO 2013/034761, WO 2012/104293 andWO2013/000924 for the treatment of neurological and psychiatricdisorders, from WO 2006/072615, WO 2006/072612, WO 2006/024640 and WO2005/113517 for the treatment of arthritis, cancer, edema and septicshock, from WO 2005/063723 for the treatment of renal and liver failure,liver dysfunction, restless leg syndrome, rheumatic disorders,arthritis, rhinitis, asthma and obesity, from WO 2005/041957 for thetreatment of cancer and thrombotic disorders, from WO 2006/102728 forthe treatment of angina pectoris and hypertension from WO 2008/043461for the treatment of cardiovascular disorders, erectile dysfunction,inflammation and renal failure and from WO 2005/061497 for the treatmentof e.g. dementia, memory disorders, cancer and osteoporosis.

Benzodiazepine like PDE2 inhibitors are described in WO 2005/063723 forthe general treatment of CNS diseases including anxiety, depression,ADHD, neurodegeneration, Alzheimer's disease and psychosis.

Newer PDE2 inhibitor families are described in WO 2015/096651, WO2015/060368 and WO 2015/012328.

Aim of the Invention

It has now been found that compounds of the present invention accordingto general formula (I) are effective inhibitors of phosphodiesterase 2.

Besides the inhibition property toward phosphodiesterase 2 enzymes, thecompounds of the present invention provide further advantageousproperties such as high selectivity with regard to PDE 10, low plasmaprotein binding across species, high CSF to plasma ratio, adequatetissue permeability and high metabolic stability.

For example the compounds of the present invention show low plasmaprotein binding across species and as a consequence high fractionunbound in plasma, high concentration in cerebrospinal fluid (CSF) andhave a high CSF to plasma ratio, which translates in lower efficaciousdoses of the compounds for disease treatment and as a consequence infurther potential advantages such as minimization of side effects.Furthermore, compounds of the present inventions show good metabolicstability both in rodents and non rodents species, good membranepermeability with no active transport at the BBB. In addition thecompounds of the present invention have very high IC50 values for PDE10.

Accordingly, one aspect of the invention refers to compounds accordingto formula (I), or salts thereof as inhibitors of phosphodiesterase 2.

Another aspect of the invention refers to compounds according to formula(I), or pharmaceutically acceptable salts thereof as inhibitors ofphosphodiesterase 2 and reaching high concentrations in cerebrospinalfluid (CSF) and/or having high CSF to plasma ratio.

Another aspect of the invention refers to compounds according to formula(I), or pharmaceutically acceptable salts thereof as inhibitors ofphosphodiesterase 2 with low plasma protein binding and thus highfraction unbound across species.

Another aspect of the invention refers to compounds according to formula(I), or pharmaceutically acceptable salts thereof as inhibitors ofphosphodiesterase 2 and showing good membrane permeability and low tomoderate in vitro efflux.

Another aspect of the invention refers to according to formula (I), orpharmaceutically acceptable salts thereof as inhibitors ofphosphodiesterase 2 and showing good metabolic stability.

In a further aspect this invention relates to pharmaceuticalcompositions, containing at least one compound according to formula (I),or pharmaceutically acceptable salts thereof, optionally together withone or more inert carriers and/or diluents.

A further aspect of the present invention relates to compounds accordingto formula (I), or pharmaceutically acceptable salts thereof orpharmaceutical compositions comprising compounds according to formula(I), or pharmaceutically acceptable salts thereof for the use in theprevention and/or treatment of disorders associated with PDE2hyperactivity and/or cAMP and/or cGMP hypofunction.

Another aspect of the invention relates to processes of manufacture ofthe compounds of the present invention.

A further aspect of the present invention relates to compounds accordingto formula (I), or pharmaceutically acceptable salts thereof orpharmaceutical compositions comprising compounds according to formula(I), or pharmaceutically acceptable salts thereof for the use in theprevention and/or treatment of diseases or conditions which can beinfluenced by inhibition of PDE2 hyperactivity and/or cAMP and/or cGMPhypofunction, such as (1) disorders comprising the symptom of cognitivedeficiency; (2) organic, including symptomatic, mental disorders,dementia; (3) mental retardation; (4) mood affective disorders; (5)neurotic, stress-related and somatoform disorders including anxietydisorders; (6) behavioural and emotional disorders with onset usuallyoccurring in childhood and adolescence, attention deficit hyperactivitysyndrome (ADHD) including Autism spectrum disorders; (7) disorders ofpsychological development, developmental disorders of scholastic skills;(8) schizophrenia and other psychotic disorders; (9) disorders of adultpersonality and behaviour; (10) mental and behavioural disorders due topsychoactive substance use; (11) extrapyramidal and movement disorders;(12) episodic and paroxysmal disorders, epilepsy; (13) Systemicatrophies primarily affecting the central nervous system, ataxia; (14)Behavioural syndromes associated with physiological disturbances andphysical factors; (15) sexual dysfunction comprising excessive sexualdrive; (16) factitious disorders; (17) obsessive-compulsive disorders;(18) depression; (19) neuropsychiatric symptoms (e.g. depressivesymptoms in Alzheimer's disease); (20) mixed dementia; (21) cognitiveimpairment in schizoaffective disorder; (22) cognitive impairment inbipolar disorder and (23) cognitive impairment in major depressivedisorder.

In addition, the compounds of the present invention can be used for thetreatment, amelioration and/or prevention of cognitive impairment beingrelated to perception, concentration, cognition, learning, attention ormemory.

In addition, the compounds of the present invention can be used for thetreatment amelioration and/or prevention of cognitive impairment beingrelated to age-associated learning and memory impairments,age-associated memory losses, vascular dementia, craniocerebral trauma,stroke, dementia occurring after strokes (post stroke dementia),post-traumatic dementia, general concentration impairments,concentration impairments in children with learning and memory problems,Alzheimer's disease, Lewy body dementia, dementia with degeneration ofthe frontal lobes, including Pick's syndrome, Parkinson's disease,progressive nuclear palsy, dementia with corticobasal degeneration,amyotropic lateral sclerosis (ALS), Huntington's disease, multiplesclerosis, thalamic degeneration, Creutzfeld-Jacob dementia, HIVdementia, schizophrenia with dementia or Korsakoff's psychosis.

In addition, the compounds of the present invention can be used for thetreatment of Alzheimer's disease.

In addition compounds of the present invention can be used for thetreatment of pain disorders, including but not limited to inflammatory,neuropathic and osteoarthritic pain.

In addition, the compounds of the present invention can be used for thetreatment of sleep disorders, bipolar disorder, metabolic syndrome,obesity, diabetes mellitus, hyperglycemia, dyslipidemia, impairedglucose tolerance, or a disease of the testes, brain, small intestine,skeletal muscle, heart, lung, thymus or spleen.

Other aims of the present invention will become apparent to the skilledman directly from the foregoing and following remarks.

DETAILED DESCRIPTION

In a first aspect the present invention relates to compounds of generalformula (I)

wherein

-   -   A is selected from the group Aa consisting of

-   -   -   wherein above mentioned groups are substituted with one R⁵            and one R⁶;

    -   R¹ is selected from the group R^(1a) consisting of        -   halogen, C₁₋₃-alkyl- and C₃₋₆-cycloalkyl-            -   wherein the above mentioned C₁₋₃-alkyl-, and                C₃₋₆-cycloalkyl-groups may optionally be substituted                with 1 to 5 substituents independently selected from the                group consisting of halogen, NC— and HO—;

    -   R² is selected from the group R^(2a) consisting of        -   aryl and heteroaryl,            -   wherein the above mentioned aryl and heteroaryl-groups                may optionally be substituted with 1 to 5 substituents                R⁴;

    -   R³ is selected from the group R^(3a) consisting of        -   H— and C₁₋₃-alkyl-,            -   wherein the above mentioned C₁₋₃-alkyl-groups may                optionally be substituted with 1 to 7 substituents                independently from each other selected from the group                consisting of halogen;

    -   R⁴ is independently from each other selected from the group        R^(4a) consisting of        -   halogen, NC—, HO—, C₁₋₄-alkyl- and C₁₋₃-alkyl-O—            -   wherein the above mentioned C₁₋₄-alkyl- and                C₁₋₃-alkyl-O-groups may optionally be substituted with 1                to 5 substituents independently selected from the group                consisting of HO— and F—;

    -   R⁵ is selected from the group R^(5a) consisting of        -   H—, halogen, NC—, HO— and C₁₋₃-alkyl-,            -   wherein the above mentioned C₁₋₃-alkyl-group may                optionally be substituted with 1 to 5 substituents                independently selected from the group consisting of HO—                and F—            -   or R⁵ and R⁶ together form an group O═;

    -   R⁶ is selected from the group R^(6a) consisting of        -   H—, halogen, NC—, HO— and C₁₋₃-alkyl-,            -   wherein the above mentioned C₁₋₃-alkyl-group may                optionally be substituted with 1 to 5 substituents                independently selected from the group consisting of HO—                and F—            -   or R⁵ and R⁶ together form a group O═;

or a salt thereof.

Unless otherwise stated, the groups, residues, and substituents,particularly R¹, R², R³, R⁴ and R⁵ are defined as above and hereinafter.If residues, substituents, or groups occur several times in a compoundthey may have the same or different meanings. Some preferred meanings ofgroups and substituents of the compounds according to the invention willbe given hereinafter.

In a further embodiment of the present invention

-   -   A is selected from the group A^(b) consisting of

-   -   -   wherein above mentioned groups are substituted with one R⁵            and one R⁶.

In a further embodiment of the present invention

-   -   A is selected from the group A⁰ consisting of

In a further embodiment of the present invention

-   -   A is selected from the group A^(d) consisting of

In a further embodiment of the present invention

-   -   A is selected from the group Ae consisting of

In a further embodiment of the present invention

-   -   R¹ is selected from the group R^(1b) consisting of F—, Cl—,        C₁₋₃-alkyl- and C₃₋₆-cycloalkyl-, wherein the above mentioned        C₁₋₃-alkyl- and C₃₋₆-cycloalkyl-groups may optionally be        substituted with 1 to 3 substituents independently selected from        the group consisting of F—.

In a further embodiment of the present invention

-   -   R¹ is selected from the group R^(1c) consisting of        -   F—, H₃C— and cyclopropyl-.

In a further embodiment of the present invention

-   -   R¹ is selected from the group R^(1d) consisting of        -   H₃C— and cyclopropyl-.

In a further embodiment of the present invention

-   -   R² is selected from the group R^(2b) consisting of        -   quinolinyl, phenyl and pyridynyl,            -   wherein the above mentioned quinoline, phenyl and                pyridyl-groups may optionally be substituted with 1 to 5                substituents R⁴.

In a further embodiment of the present invention

-   -   R² is selected from the group R^(2c) consisting of        -   phenyl and pyridyl,            -   wherein the above mentioned phenyl and pyridyl-groups                may optionally be substituted with 1 to 2 substituents                R⁴.

In a further embodiment of the present invention

-   -   R² is selected from the group R^(2d) being

In a further embodiment of the present invention

-   -   R² is selected from the group R^(2e) being

In a further embodiment of the present invention

-   -   R² is selected from the group R^(2f) being

In a further embodiment of the present invention

-   -   R³ is selected from the group R^(3b) consisting of        -   H—, H₃C—, F₃C—, F₂HC—, FH₂C— and F₃C—.

In a further embodiment of the present invention

-   -   R³ is selected from the group R^(3c) consisting of    -   H— and H₃C—.

In a further embodiment of the present invention

-   -   R³ is selected from the group R^(3d) being H—.

In a further embodiment of the present invention

-   -   R⁴ is independently from each other selected from the group        R^(4b) consisting of    -   halogen, C₁₋₄-alkyl- and C₁₋₃-alkyl-O—        -   wherein the above mentioned C₁₋₄-alkyl- and            C₁₋₃-alkyl-O-groups may optionally be substituted with 1 to            5 substituents independently selected from the group            consisting of HO—, and F—.

In a further embodiment of the present invention

-   -   R⁴ is independently from each other selected from the group        R^(4c) consisting of        -   halogen, C₁₋₃-alkyl-, F₃C—O—, F₂HC—O—, FH₂C—O— and H₃C—O—,            -   wherein the above mentioned C₁₋₃-alkyl-groups may                optionally be substituted with 1 to 5 F—.

In a further embodiment of the present invention

-   -   R⁴ is independently from each other selected from the group        R^(4d) consisting of        -   F, Cl, Br, F₃C—, F₂HC—, FH₂C—, H₃C—, F₃C—O—, F₂HC—O—,            FH₂C—O— and H₃C—O—.

In a further embodiment of the present invention

-   -   R⁴ is independently from each other selected from the group        R^(4e) consisting of        -   F, C₁, F₃C—, F₃C—O— and H₃C—O—.

In a further embodiment of the present invention

-   -   R⁴ is independently from each other selected from the group        R^(4f) consisting of        -   F and F₃C—.

In a further embodiment of the present invention

-   -   R⁵ is selected from the group R^(5b) consisting of        -   H—, HO— and C₁₋₂-alkyl-,            -   wherein the above mentioned C₁₋₂-alkyl-group may                optionally be substituted with 1 to 5 F—, or R⁵ and R⁶                together form an group O═.

In a further embodiment of the present invention

-   -   R⁵ is selected from the group R^(5c) consisting of        -   H— and HO—.

In a further embodiment of the present invention

-   -   R⁵ is selected from the group R^(5d) being        -   HO—.

In a further embodiment of the present invention

-   -   R⁶ is selected from the group R^(6b) consisting of        -   H— and C₁₋₂-alkyl-,            -   wherein the above mentioned C₁₋₂-alkyl-group may                optionally be substituted with 1 to 5 F—, or R⁵/R⁶                together form a group O═.

In a further embodiment of the present invention

-   -   R⁶ is selected from the group R^(6c) consisting of        -   H and H₃C—,            -   wherein the above mentioned methyl-group may optionally                be substituted with 1 to 3 F—.

In a further embodiment of the present invention

-   -   R⁶ is selected from the group R^(6d) consisting of        -   H— and H₃C—.

Each A^(x), R^(1x), R^(2x), R^(3x), R^(4x), R^(5x) and R^(6x) representsa characterized, individual embodiment for the corresponding substituentas described above. Thus given the above definitions, individualembodiments of the first aspect of the invention are fully characterizedby the term (A^(x), R^(1x), R^(2x), R^(3x), R^(4x), R^(5x) and R^(6x)),wherein for each index x an individual figure is given that ranges from“a” to the highest letter given above. All individual embodimentsdescribed by the term in parentheses with full permutation of theindices x, referring to the definitions above, shall be comprised by thepresent invention.

The following Table 1 shows such embodiments E-1 to E-39 of theinvention that are considered preferred. Embodiment E-39, represented bythe entries in the last row of Table 1, is the most preferredembodiment.

TABLE 1 Embodiments E-1 to E-39 of the invention A^(x) R^(1x) R^(2x)R^(3x) R^(4x) R^(5x) R^(6x) E-1 A^(a) R^(1a) R^(2a) R^(3a) R^(4b) R^(5a)R^(6a) E-2 A^(a) R^(1a) R^(2a) R^(3b) R^(4b) R^(5a) R^(6a) E-3 A^(a)R^(1b) R^(2b) R^(3b) R^(4c) R^(5a) R^(6a) E-4 A^(a) R^(1c) R^(2b) R^(3b)R^(4d) R^(5a) R^(6a) E-5 A^(a) R^(1c) R^(2b) R^(3c) R^(4e) R^(5b) R^(6b)E-6 A^(b) R^(1b) R^(2b) R^(3b) R^(4b) R^(5a) R^(6a) E-7 A^(b) R^(1c)R^(2b) R^(3c) R^(4e) R^(5b) R^(6b) E-8 A^(c) R^(1c) R^(2b) R^(3b) R^(4b)R^(5b) R^(6b) E-9 A^(c) R^(1c) R^(2b) R^(3c) R^(4e) R^(5b) R^(6b) E-10A^(c) R^(1c) R^(2c) R^(3b) R^(4c) R^(5b) R^(6b) E-11 A^(c) R^(1d) R^(2c)R^(3c) R^(4d) R^(5b) R^(6b) E-12 A^(c) R^(1d) R^(2c) R^(3d) R^(4e)R^(5b) R^(6b) E-13 A^(c) R^(1d) R^(2c) R^(3d) R^(4f) R^(5c) R^(6c) E-14A^(c) R^(1d) R^(2d) R^(3b) R^(4c) R^(5b) R^(6b) E-15 A^(c) R^(1d) R^(2d)R^(3c) R^(4d) R^(5b) R^(6b) E-16 A^(c) R^(1d) R^(2d) R^(3c) R^(4f)R^(5d) R^(6d) E-17 A^(c) R^(1d) R^(2d) R^(3d) R^(4e) R^(5b) R^(6b) E-18A^(c) R^(1d) R^(2d) R^(3d) R^(4f) R^(5c) R^(6c) E-19 A^(c) R^(1d) R^(2e)R^(3b) R^(4b) R^(5b) R^(6b) E-20 A^(c) R^(1d) R^(2e) R^(3b) R^(4b)R^(5c) R^(6c) E-21 A^(c) R^(1d) R^(2e) R^(3c) R^(4d) R^(5b) R^(6b) E-22A^(c) R^(1d) R^(2e) R^(3c) R^(4d) R^(5c) R^(6c) E-23 A^(c) R^(1d) R^(2e)R^(3c) R^(4e) R^(5b) R^(6b) E-24 A^(c) R^(1d) R^(2e) R^(3c) R^(4e)R^(5c) R^(6c) E-25 A^(c) R^(1d) R^(2e) R^(3d) R^(4e) R^(5b) R^(6b) E-26A^(c) R^(1d) R^(2e) R^(3d) R^(4e) R^(5c) R^(6c) E-27 A^(c) R^(1d) R^(2f)R^(3c) ^(—) R^(5d) R^(6d) E-28 A^(c) R^(1d) R^(2f) R^(3d) ^(—) R^(5d)R^(6d) E-29 A^(d) R^(1c) R^(2c) R^(3b) R^(4d) R^(5b) R^(6b) E-30 A^(d)R^(1c) R^(2d) R^(3b) R^(4e) R^(5c) R^(6c) E-31 A^(d) R^(1c) R^(2e)R^(3c) R^(4e) R^(5d) R^(6d) E-32 A^(d) R^(1d) R^(2f) R^(3c) ^(—) R^(5d)R^(6d) E-33 A^(e) R^(1c) R^(2c) R^(3b) R^(4d) R^(5b) R^(6b) E-34 A^(e)R^(1c) R^(2d) R^(3b) R^(4e) R^(5c) R^(6c) E-35 A^(e) R^(1c) R^(2e)R^(3c) R^(4e) R^(5d) R^(6d) E-36 A^(e) R^(1c) R^(2f) R^(3b) ^(—) R^(5b)R^(6b) E-37 A^(e) R^(1d) R^(2f) R^(3c) ^(—) R^(5c) R^(6c) E-38 A^(e)R^(1d) R^(2f) R^(3c) ^(—) R^(5d) R^(6d) E-39 A^(e) R^(1d) R^(2f) R^(3d)^(—) R^(5d) R^(6d)

Accordingly, for example E-1 covers compounds of formula (I), wherein

-   -   A is selected from the group Aa consisting of

-   -   -   wherein above mentioned groups are substituted with one R⁵            and one R⁶;

    -   R¹ is selected from the group R^(1a) consisting of        -   halogen, C₁₋₃-alkyl- and C₃₋₆-cycloalkyl-            -   wherein the above mentioned C₁₋₃-alkyl-, and                C₃₋₆-cycloalkyl-groups may optionally be substituted                with 1 to 5 substituents independently selected from the                group consisting of halogen, NC— and HO—;

    -   R² is selected from the group R^(2a) consisting of        -   aryl and heteroaryl,            -   wherein the above mentioned aryl and heteroaryl-groups                may optionally be substituted with 1 to 5 substituents                R⁴;

    -   R³ is selected from the group R^(3a) consisting of        -   H— and C₁₋₃-alkyl-,            -   wherein the above mentioned C₁₋₃-alkyl-groups may                optionally be substituted with 1 to 7 substituents                independently from each other selected from the group                consisting of halogen;

    -   R⁴ is independently from each other selected from the group        R^(4b) consisting of        -   halogen, C₁₋₄-alkyl- and C₁₋₃-alkyl-O—            -   wherein the above mentioned C₁₋₄-alkyl- and                C₁₋₃-alkyl-O-groups may optionally be substituted with 1                to 5 substituents independently selected from the group                consisting of HO—, and F—;

    -   R⁵ is selected from the group R^(5a) consisting of        -   H—, halogen, NC—, HO— and C₁₋₃-alkyl-,            -   wherein the above mentioned C₁₋₃-alkyl-group may                optionally be substituted with 1 to 5 substituents                independently selected from the group consisting of HO—                and F—            -   or R⁵ and R⁶ together form an group O═;

    -   R⁶ is selected from the group R^(6a) consisting of        -   H—, halogen, NC—, HO— and C₁₋₃-alkyl-,            -   wherein the above mentioned C₁₋₃-alkyl-group may                optionally be substituted with 1 to 5 substituents                independently selected from the group consisting of HO—                and F—            -   or R⁵ and R⁶ together form a group O═;

or a salt thereof.

Accordingly, for example E-5 covers compounds of formula (I), wherein

-   -   A is selected from the group Aa consisting of

-   -   -   wherein above mentioned groups are substituted with one R⁵            and one R⁶;

    -   R² is selected from the group R^(2b) consisting of        -   quinolinyl, phenyl and pyridynyl,            -   wherein the above mentioned quinoline, phenyl and                pyridyl-groups may optionally be substituted with 1 to 5                substituents R⁴;

    -   R³ is selected from the group R^(3c) consisting of        -   H— and H₃C—;

    -   R⁴ is independently from each other selected from the group        R^(4e) consisting of        -   F, C₁, F₃C—, F₃C—O— and H₃C—O—;

    -   R⁵ is selected from the group R^(5b) consisting of        -   H—, HO— and C₁₋₂-alkyl-,            -   wherein the above mentioned C₁₋₂-alkyl-group may                optionally be substituted with 1 to 5 F—, or R⁵ and R⁶                together form an group O═:

    -   R⁶ is selected from the group R^(6b) consisting of        -   H— and C₁₋₂-alkyl-,            -   wherein the above mentioned C₁₋₂-alkyl-group may                optionally be substituted with 1 to 5 F—, or R⁵/R⁶                together form a group O═;

or a salt thereof.

Accordingly, for example E-39 covers compounds of formula (I), wherein

-   -   A is selected from the group Ae consisting of

-   -   R¹ is selected from the group R^(1d) consisting of        -   H₃C— and cyclopropyl-;    -   R² is selected from the group R^(2f) being

R³ is selected from the group R^(3d) being H—;

-   -   R⁵ is selected from the group R^(5d) being        -   HO—;    -   R⁶ is selected from the group R^(6d) consisting of        -   H— and methyl-;

or a salt thereof.

Further preferred are the following compounds listed in Table 2:

No. Structure I

II

III

IV

V

VI

VII

VIII

IX

X

XI

XII

XIII

XIV

XV

XVI

XVII

XVIII

XIX

XX

XXI

XXII

XXIII

XXIV

XXV

XXVI

XXVII

XXVIII

XXIX

XXX

XXXI

XXXII

XXXIII

XXXIV

XXXV

XXXVI

XXXVII

XXXVIII

XXXIX

XL

XLI

XLII

XLIII

XLIV

XLV

XLVI

XLVII

XLVIII

XLIX

L

LI

LII

LIII

LIV

LV

LVI

LVII

LVIII

LIX

LX

LXI

LXII

LXIII

LXIV

LXV

LXVI

LXVII

LXVIII

LXIX

LXX

LXXI

LXXII

LXXIII

LXXIV

LXXV

LXXVI

LXXVII

or the salts thereof.

Some terms used above and hereinafter to describe the compoundsaccording to the invention will now be defined more closely.

Terms not specifically defined herein should be given the meanings thatwould be given to them by one of skill in the art in light of thedisclosure and the context. As used in the specification, however,unless specified to the contrary, the following terms have the meaningindicated and the following conventions are adhered to. In the groups,radicals, or moieties defined below, the number of carbon atoms is oftenspecified preceding the group, for example C₁₋₆-alkyl means an alkylgroup or radical having 1 to 6 carbon atoms. In general, for groupscomprising two or more subgroups, the last named subgroup is the radicalattachment point, for example, the substituent “aryl-C₁₋₃-alkyl-” meansan aryl group which is bound to a C₁₋₃-alkyl-group, the latter of whichis bound to the core molecule or to the group to which the substituentis attached.

Within the present invention, the term “core molecule” is defined by thefollowing structure:

In general, the attachment site of a given residue to another groupshall be variable, i.e. any capable atom, bearing hydrogens to bereplaced, within this residue may be the linking spot to the group beingattached, unless otherwise indicated.

In case a compound of the present invention is depicted in form of achemical name and as a formula in case of any discrepancy the formulashall prevail.

An asterisk may be used in sub-formulas to indicate the bond orattachment point which is connected to the core molecule, rest of themolecule or to the substituent to which it is bound as defined.

Unless specifically indicated, throughout the specification and theappended claims, a given chemical formula or name shall encompasstautomers and all stereo, optical and geometrical isomers (e.g.enantiomers, diastereomers, E/Z isomers etc. . . . ) and racematesthereof as well as mixtures in different proportions of the separateenantiomers, mixtures of diastereomers, or mixtures of any of theforegoing forms where such isomers and enantiomers exist, as well assalts, including pharmaceutically acceptable salts thereof and solvatesthereof such as for instance hydrates including solvates of the freecompounds or solvates of a salt of the compound.

The phrase “pharmaceutically acceptable” or “physiologically acceptable”is employed herein to refer to those compounds, materials, compositions,and/or dosage forms which are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of human beingsand animals without excessive toxicity, irritation, allergic response,or other problem or complication, and commensurate with a reasonablebenefit/risk ratio.

As used herein, “pharmaceutically acceptable salts” or “physiologicallyacceptable salts” refer to derivatives of the disclosed compoundswherein the parent compound is modified by making acid or base saltsthereof. Examples of pharmaceutically acceptable salts orphysiologically acceptable salts include, but are not limited to,mineral or organic acid salts of basic residues such as amines; alkalior organic salts of acidic residues such as carboxylic acids; and thelike. For example, such salts include salts from ammonia, L-arginine,betaine, benethamine, benzathine, calcium hydroxide, choline, deanol,diethanolamine (2,2′-iminobis(ethanol)), diethylamine,2-(diethylamino)-ethanol, 2-aminoethanol, ethylenediamine,N-ethyl-glucamine, hydrabamine, 1H-imidazole, lysine, magnesiumhydroxide, 4-(2-hydroxyethyl)-morpholine, piperazine, potassiumhydroxide, 1-(2-hydroxyethyl)-pyrrolidine, sodium hydroxide,triethanolamine (2,2′,2″-nitrilotris(ethanol)), tromethamine, zinchydroxide, acetic acid, 2,2-dichloro-acetic acid, adipic acid, alginicacid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoicacid, 2,5-dihydroxybenzoic acid, 4-acetamido-benzoic acid, (+)-camphoricacid, (+)-camphor-10-sulfonic acid, carbonic acid, cinnamic acid, citricacid, cyclamic acid, decanoic acid, dodecylsulfuric acid,ethane-1,2-disulfonic acid, ethanesulfonic acid,2-hydroxy-ethanesulfonic acid, ethylenediaminetetraacetic acid, formicacid, fumaric acid, galactaric acid, gentisic acid, D-glucoheptonicacid, D-gluconic acid, D-glucuronic acid, glutamic acid, glutaric acid,2-oxo-glutaric acid, glycerophosphoric acid, glycine, glycolic acid,hexanoic acid, hippuric acid, hydrobromic acid, hydrochloric acid,isobutyric acid, DL-lactic acid, lactobionic acid, lauric acid, lysine,maleic acid, (−)-L-malic acid, malonic acid, DL-mandelic acid,methanesulfonic acid, galactaric acid, naphthalene-1,5-disulfonic acid,naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid,nitric acid, octanoic acid, oleic acid, orotic acid, oxalic acid,palmitic acid, pamoic acid (embonic acid), phosphoric acid, propionicacid, (−)-L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid,sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid,(+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid andundecylenic acid. Further pharmaceutically acceptable salts can beformed with cations from metals like aluminium, calcium, lithium,magnesium, potassium, sodium, zinc and the like (also see Pharmaceuticalsalts, Berge, S. M. et al., J. Pharm. Sci., (1977), 66, 1-19).

The pharmaceutically acceptable salts of the present invention can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha sufficient amount of the appropriate base or acid in water or in anorganic diluent like ether, ethyl acetate, ethanol, isopropanol, oracetonitrile, or a mixture thereof.

Salts of other acids than those mentioned above which for example areuseful for purifying or isolating the compounds of the present invention(e.g. trifluoro acetate salts) also comprise a part of the invention.

The term “substituted” as used herein means that any one or morehydrogens on the designated atom is replaced with a selection from theindicated group, provided that the designated atom's viable valencenumber is not exceeded, and that the substitution results in a stablecompound.

The term “partially unsaturated” as used herein means that in thedesignated group or moiety 1, 2, or more, preferably 1 or 2, doublebonds are present. Preferably, as used herein, the term “partiallyunsaturated” does not cover fully unsaturated groups or moieties.

The term “halogen” generally denotes fluorine (F), chlorine (Cl),bromine (Br) and iodine (1).

The term “C₁-n-alkyl”, wherein n is an integer from 2 to n, either aloneor in combination with another radical denotes an acyclic, saturated,branched or linear hydrocarbon radical with 1 to n C atoms. For examplethe term C₁₋₅-alkyl embraces the radicals H₃C—, H₃C—CH₂—, H₃C—CH₂—CH₂—,H₃C—CH(CH₃)—, H₃C—CH₂—CH₂—CH₂—, H₃C—CH₂—CH(CH₃)—, H₃C—CH(CH₃)—CH₂—,H₃C—C(CH₃)₂—, H₃C—CH₂—CH₂—CH₂—CH₂—, H₃C—CH₂—CH₂—CH(CH₃)—,H₃C—CH₂—CH(CH₃)—CH₂—, H₃C—CH(CH₃)—CH₂—CH₂—, H₃C—CH₂—C(CH₃)₂—,H₃C—C(CH₃)₂—CH₂—, H₃C—CH(CH₃)—CH(CH₃)— and H₃C—CH₂—CH(CH₂CH₃)—.

The term “C₃-n-cycloalkyl”, wherein n is an integer from 4 to n, eitheralone or in combination with another radical denotes a cyclic,saturated, unbranched hydrocarbon radical with 3 to n C atoms. Forexample the term C₃₋₇-cycloalkyl includes cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and cycloheptyl.

The term “aryl” as used herein, either alone or in combination withanother radical, denotes a carbocyclic aromatic monocyclic groupcontaining 6 carbon atoms which may be further fused to a second 5- or6-membered carbocyclic group which may be aromatic, saturated orunsaturated. Aryl includes, but is not limited to, phenyl, indanyl,indenyl, naphthyl, anthracenyl, phenanthrenyl, tetrahydronaphthyl anddihydronaphthyl.

The term “heteroaryl” means a mono- or polycyclic-ring systemscontaining one or more heteroatoms selected from N, O or S(O)r, whereinr=0, 1 or 2, consisting of 5 to 14 ring atoms wherein at least one ofthe heteroatoms is part of an aromatic ring. The term “heteroaryl” isintended to include all the possible isomeric forms.

In one embodiment the term “heteroaryl” means a mono- or bicyclic-ringsystem containing one to three heteroatoms selected from N, O or S(O)r,wherein r=0, 1 or 2, consisting of 5 to 10 ring atoms wherein at leastone of the heteroatoms is part of an aromatic ring.

Thus, the term “heteroaryl” includes the following exemplary structureswhich are not depicted as radicals as each form may be attached througha covalent bond to any atom so long as appropriate valences aremaintained:

Many of the terms given above may be used repeatedly in the definitionof a formula or group and in each case have one of the meanings givenabove, independently of one another.

The compounds according to the invention may be obtained using methodsof synthesis known in principle. Preferably, the compounds are obtainedby the following methods according to the invention which are describedin more detail hereinafter.

Preparation

The following Schemes shall illustrate generally how to manufacture thecompounds of the present invention by way of example. The abbreviatedsubstituents may be as defined above if not defined otherwise within thecontext of the schemes.

The preparation process might comprises:

-   -   a) Reacting a compound of formula (II)

or a derivatives thereof, with a compound of formula (III)

Wherein R¹, R², R³, R⁵, R⁶ and A are as defined above and L is asuitable leaving group such as halogen atom (e.g. chlorine or bromine)or hydroxyl group.

In case of L=halogen, process a) typically comprises the reaction of acompound of formula (II) with a compound of formula (III) in anappropriate solvent such as acetonitrile or N,N-dimethylformamide in thepresence of a base such as TEA or DIPEA at room temperature.

In case of L=OH, process a) typically comprises the reaction of acompound of formula (II) with a compound of formula (III) in anappropriate solvent such as N,N-dimethylformamide and in the presence ofa suitable coupling agent (e.g. HATU or TBTU)

Compounds of formula (III) are either commercially available or can beprepared as described in the following Schemes, following known reportedprocedures.

In Scheme 1, Step 1 typically involves reaction of commerciallyavailable amino pyrazole derivatives with 2-Bromo-malonaldehyde in thepresence of acetic acid in a suitable solvent such as EtOH underheating. In Step 2, the cyclopropyl group is introduced by a crosscoupling palladium catalyzed reaction using for example potassiumcyclopropyltrifluoroborate, a suitable palladium catalyst such asPalladium(II) acetate and 2-dicyclohexylphosphino-2′,6′-diisopropoxy1,1′-biphenyl as ligand in an appropriate solvent such as toluene underheating. In Step 3 the ethyl ester is then hydrolyzed under basicconditions using sodium hydroxide or lithium hydroxide monohydrate in anappropriate solvent such as EtOH or a mixture of THE/water.

In Scheme 2, Step 1 typically involves reaction of commerciallyavailable amino pyrazole derivative with 1,1,3,3,-tetraethoxy-propane inthe presence of hydrochloric acid in a suitable solvent such as EtOHunder heating. Bromination using bromine in acetic acid as solvent atroom temperature provides the bromo derivative and the cyclopropyl groupis then introduced as described in Scheme 1.

In Scheme 3, Step 1 typically involves reaction of commerciallyavailable amino pyrazole derivative with1,1,3,3,-tetraethoxy-2-methyl-propane in the presence of hydrochloricacid in a suitable solvent such as EtOH under heating. Basic hydrolysisprovides the desired carboxylic acid derivative

Compounds of formula (II) are either commercially available or can beprepared as described in the following Schemes.

In Scheme 4, R² is aryl or heteroaryl

In Scheme 4, Step 1, commercially available ketone is converted in thecorresponding 2-methyl-propane-2-sulfinyl-imine using titanium (IV)ethoxide and 2-Methyl-propane-2-sulfinic acid amide, as described in WO2005087751.

The obtained intermediate is then added dropwise to a previouslyprepared solution of organo lithium derivatives of the appropriatehalogen compounds (R²X, where X is bromine or iodine) prepared using forexample commercially available solution of tert-butyllithium orn-buthyllitium in hexane or pentane at low temperature (−75° C.) in asuitable solvent such as toluene or THF. Cleavage of the resultingsulphinic amide by treatment with acid such as a 4N solution of HCl in asuitable solvent such as dioxane provides the desired intermediatesamines.

The above described synthesis applies also for the analogues with 5 and4 membered ring, starting from commercially available cyclopentanone andoxetane-3-one.

In Scheme 5, R² is aryl derivatives.

Step 1 involves a cross coupling Suzuky reaction with commerciallyavailable boronic acid or pinacol ester derivatives and the appropriatehalogen derivatives (X═Br or I) using for example1,1′-bis(diphenylphosphino)ferrocenedichloro palladium(II) as catalyst,potassium carbonate as base in an appropriate solvent such as a mixtureof toluene/water under heating. The epoxidation step is performed usingMCPBA as oxidating agent in DCM at room temperature. The desired aminoalcohol intermediate is then obtained by opening the epoxide with amodified Ritter procedure using trifluoromethane sulfonic acid andacetonitrile followed by basic hydrolysis of the formed intermediate, inanalogy to the procedure described in Tetrahedron Asymmetry, 1996, 5,1501-1506.

The relative stereochemistry of the above described aminoalcohols isreported in the Experimental description.

In Scheme 6, R² is aryl or heteroaryl.

In Scheme 6, the opening of the epoxide is performed using sodium azidein the presence of ammonium chloride under heating in a suitablesolvent, such as dimethyl formamide. After separation of the tworegioisomers, (see experimental), the azide group is then converted intoamino group by reduction following well known reported procedure such asfor example using zinc and ammonium formate in a suitable solvent suchas methanol at room temperature.

The relative stereochemistry of the above described amino alcohols isreported in the Experimental description.

In Scheme 7, R² is aryl or heteroaryl

In Scheme 7, the regioisomeric mixture of azide intermediates, obtainedfollowing the approach described in Scheme 6, is reduced under catalytichydrogenation conditions, using for example Pd/C in a suitable solventsuch as ethanol and in the presence of di.tert-butyldicarbonate toobtain the protected amino alcohols derivatives. Oxidation to ketone isperformed using Dess Martin periodinane in a suitable solvent such asDCM at room temperature or using Swern's procedure Formation of thetertiary alcohols is accomplished by addition of methyl magnesiumchloride to the carbonyl group at low temperature (−20° C.) in asuitable solvent such as THF. The cleavage of the Boc protecting groupis performed under acid conditions using for example trifluoroaceticacid in a suitable solvent such as DCM at room temperature.

The regioisomeric ratio of epoxide opening and the relativestereochemistry of the above described amino alcohols are reported inthe Experimental description.

In Scheme 8 R² is aryl.

In Scheme 8, the desired tetrahydrofuran-3-ol-intermediates are obtainedby addition of the appropriate lithium derivatives, prepared reacting asuitable halogen compounds (R²X, X=halogen) with commercially availablesolution of tert-butyllithium or n-buthyllitium in hexane or pentane atlow temperature (−75° C.) in a suitable solvent such as toluene or THF,to the carbonyl group. Treatment with pTsOH in toluene under refluxprovides the double bond derivatives which are transformed into thedesired aminoalcohols following the approaches described in Scheme 5.

The relative stereochemistry of the aminoalcohols compounds are reportedin the Experimental description.

In Scheme 9, R² is aryl or heteroaryl.

The desired amino alcohols are obtained following the approachesdescribed above in Scheme 6 and 7.

The regiochemistry ratio of the epoxide opening and the relativestereochemistry of the aminoalcohols compounds are reported in theExperimental description.

Biological Examples

In-Vitro Effect:

The in-vitro effect of the active compounds of the invention can beshown with the following biological assays.

a) Phosphodiesterase (PDE) 2A and 10 Assay with Fluorescent Substrate

Assay Principle:

The PDE reaction cleaves cAMP to AMP. The IMAP system (Molecular Device)using fluorescence polarization (FP) as detection principle was used tomeasure enzyme activity. A fluorescent labeled cAMP was used assubstrate for the reaction, generating a labeled AMP. The fluorescentAMP binds specifically to the large M(Ill)-based nano-particles whichreduces the rotational speed of the substrate and thus increases itspolarization.

Detailed Method:

The inhibition of PDE 2A or 10 enzyme activity was assessed usingIMAP-Phosphodiesterase-cAMP fluorescence labeled substrate (MolecularDevices, Order No. R7506), IMAP TR-FRET screening express (MolecularDevices, Order No. R8160, the TR-FRET component will not be used) andPDE 2A or PDE10 protein expressed upon baculovirus infection in SF9cells. The cells were incubated after infection for ˜3 days and proteinproduction was confirmed by Western Blot. The cells were collected bycentrifugation and the pellet frozen in liquid nitrogen before it wasresuspended in PBS containing 1% Triton X-100 and protease inhibitors.After 45 min incubation on ice, the cell debris was removed bycentrifugation (13.000 rpm, 30 min). Since SF 9 cells do not expresscAMP hydrolyzing enzymes to a high extent, no further purification ofthe protein was needed.

All reactions were performed in 384 well plates, Perkin Elmer blackoptiplates and IMAP reaction buffer with 0.1% Tween20 (kit component)

Compounds were serial diluted in DMSO. With an intermediate dilutionstep with reaction buffer DMSO concentration was reduced to achieve 1%DMSO in the assay reaction. Setup of the assay started with 10 μl enzyme(˜1 Ong/well, depending on prep. batch), 5 μl compound, reaction wasstarted by addition of 5 μl labeled cAMP (30 nM, final concentration),immediately mixed for 15 seconds on a Eppendorf mixmate (2000 rpm)followed by an incubation at room temperature for 90 minutes in thedark. Reaction is stopped by adding of 60 μl binding buffer for FP/cAMP(kit component). After at least 90 min of further incubation (roomtemperature, dark) the assay was measured at 485 nm excitation/525 nmemission in an Envision multilabel reader (PerkinElmer).

Each assay plate contained wells with vehicle controls (1% DMSO) for themeasurement of non-inhibited reaction (=100% control) and wells withoutenzyme as 0% controls.

The analysis of the data was performed by calculation of the percentageof inhibition in the presence of test compound compared to the vehiclecontrol samples (100% control, no inhibition) and a low control (0%control, no enzyme).

IC50 values are calculated with Assay Explorer or other suited softwarebased on curve fitting of results of at least 8 different compoundconcentrations. The compound concentrations may vary according to theneeded range, but typically cover the range between 10 μM and 0.1 pM.

TABLE 3a PDE2A Activity of the examples (Ex) compiled in theexperimental part, based on above described assay (IMAP fluorescent).PDE2 Ex. IC₅₀ [nM]   1 14   2 22   3 266   4 48   5 258   6 199   7 77  8 69   9 117  10 80  11 297  12 1650  13 359  14 456  15 746  16 537 17 39  18 129  19 519  20 172  21 74  22 119  23 232  24 754  25 88  26174  27 534  28 834  29 661  30 12  31 30  32 67  33 83  34 27  35 31 36 180  37 192  38 331  39 568  40 84  41 291  42 130  43 359  44 840 45 239  46 5.7  47 240  48 1.35  49 70  50 596  51 59  52 231  53 100 54 12  55 21  56 229  57 103  58 14  59 60  60 22  61 19  62 127  63124  64 496  65 30  66 25  67 50  68 24  69 1740  70 1250  71 13  72 122 73 1142  74 2530  75 42 80b 75 80a 313 81a 3.4 81b 149 82a 1.2 82b 4083a 5.9 83b 1820 84a 120 84b 2790 85a 130 86a 3.9 86b 2550 87a 1000 87b184 88a 32 88b 3030 89a 1000 90a 200 90b 14 91a 63 91b 1590 92a 93 92b712 93a 11 93b 1520

TABLE 3b PDE10 Activity of the examples (Ex) compiled in theexperimental part, based on above described assay (IMAP fluorescent).PDE10 Ex. IC₅₀ [nM]   1 10100   2 >10000   3 >10000   4 550   5 12200  6 >10000   7 9110   8 >10000   9 >10000  10 9820  11 1470  12 >10000 13 9910  14 8430  15 >10000  16 >10000  17 6940  18 8630  19 >10000 20 >10000  21 >10000  22 9920  23 >10000  24 >10000  25 9070  26 >10000 27 >10000  28 >10000  29 5930  30 >10000  31 >10000  32 >10000 33 >10000  34 10800  35 >10000  36 >10000  37 >10000  38 6710 39 >10000  40 5730  41 7950  42 5590  43 6860  44 9680  45 7850 46 >10000  47 >10000  48 6620  49 >10000  50 >10000  51 9040  52 >10000 53 >10000  54 9670  55 >10000  56 >10000  57 >10000  58 >10000 59 >10000  60 >10000  61 6650  62 7160  63 >10000  64 >10000  65 9760 66 >10000  67 >10000  68 >10000  69 >10000  70 >10000  71 >10000 72 >10000  73 >10000  74 >10000  75 >10000 80b 5280 80a 7760 81a >1000081b >10000 82a >10000 82b 8461 83a >10000 83b >10000 84a >1000084b >10000 85a >10000 86a >10000 86b 9940 87a >10000 87b >10000 88a 756088b >10000 89a >10000 90a 8590 90b 7350 91a 7700 91b 5670 92a >1000092b >10000 93a >10000 93b >10000

In-Vivo Effect:

Animal Experiments and Sample Analysis (CSF):

Test compounds were administered to animals (rat) different routes atdoses of 10.0 or 5 μmol/kg, (both oral and intravenous). CSF sampleswere carefully collected by puncture of the cisterna magna underanesthaesia. Immediately after CSF sampling, blood was taken by heartpuncture and brains were dissected out. Blood was collected inEDTA-coated microvettes and plasma was prepared by centrifugation.Concentration of the test compounds in plasma, CSF or brain homogenatewas determined using HPLC-MS-MS.

TABLE 4 Plasma, brain and CSF concentration conc conc conc Time(*)plasma brain c(brain)/ CSF c(CSF)/ Ex. (h) (nmol/L) (nmol/L) c(plasma)(nmol/L) c(plasma)   1 0.5 243 471 1.96 11 0.04  21 0.5 1210 1320 1.17106 0.09  25 0.5 1040 957 0.92 111 0.12 81a 0.5 2460 1070 0.42 261 0.1082a 0.5 3320 1180 0.36 157 0.05 83a 0.5 794 449 0.6 61 0.08 (*)Timebetween administration and CSF sampling

For the skilled in the art it is evident from the experimental resultsshown above that compounds of the present invention are not only potentphosphodiesterase 2 inhibitors but also reach high CSF concentrationsand adequate CSF to plasma ratios.

Plasma Protein Binding (Determination of Human and Rat Plasma ProteinBinding with Equilibrium Dialysis)

This equilibrium dialysis (ED) technique is used to determine theapproximate in vitro fractional binding of test compounds to human andrat plasma proteins.

Dianorm Teflon dialysis cells (micro 0.2) are used. Each cell consistsof a donor and an acceptor chamber, separated by an ultrathinsemipermeable membrane with a 5 kDa molecular weight cutoff.

Stock solutions for each test compound are prepared in DMSO at 1 mM anddiluted to a final concentration of 1.0 μM. The subsequent dialysissolutions are prepared in pooled human and rat plasma (with NaEDTA)

Aliquots of 200 μL dialysis buffer (100 mM potassium phosphate, pH 7.4)are dispensed into the buffer chamber. Aliquots of 200 μL test compounddialysis solution are dispensed into the plasma chambers. Incubation iscarried out for 2 hours under rotation at 37° C.

At the end of the dialysis period, the dialysate is transferred intoreaction tubes. The tubes for the buffer fraction contain 0.2 mlAcetonitril/water (80/20). Aliquots of 25 μL of the plasma dialysate aretransferred into deep well plates and mixed with 25 μl Acetonitril/water(80/20), 25 μl buffer, 25 μL calibration solution and 25 μl InternalStandard solution. Protein precipitation is done by adding 200 μlAcetonitrile. Aliquots of 50 μl of the buffer dialysate are transferredinto deep well plates and mixed with 25 μl blank plasma, 25 μl InternalStandard solution and 200 μl Acetonitril. Samples are measured onHPLC-MS/MS-Systems and evaluated with Analyst-Software.

Percent bound is calculated with the formula: % bound=(plasmaconcentration−buffer concentration/plasma concentration)×100 and % freeis calculated as difference.

TABLE 4 PPB (Plasma Protein Binding) of compounds of the presentinvention in human and rat plasma. PPB HUM PPB RAT EX % BINDING %BINDING  2 91.5 94.7  1 96 96.8 31 95.3 97.6 35 92.4 93.7 37 94.4 93.5017 91.4 90.7 25 75.4 83.1 21 83.8 87.2  3 84.3 — 68 84.4 — 69 90.7 — 7079.4 — 71 81.1 — 51 46.9 — 53 81.7 — 88b 44.2 — 82a 78.7 86.5 81a 63.074.3 46 67.1 68.7 81b 65.3 66.2 83a 84.4 82.2 86a 89.5 93.5

For the skilled in the art it is evident from the experimental resultsshown above that compounds of the present invention are not only potentphosphodiesterase 2 inhibitors but also have low plasma protein binding.

Assessment of Efflux in Madin-Darby Canine Kidney Cells Transfected withthe Human MDR1 Gene (MDCK Assay)

Apparent permeability coefficients (PE) of the compounds across theMDCK-MDR1 cell monolayers are measured (pH 7.4, 37° C.) inapical-to-basal (AB) and basal-to-apical (BA) transport direction. ABpermeability (PEAB) represents drug absorption from the blood into thebrain and BA permeability (PEBA) drug efflux from the brain back intothe blood via both passive permeability as well as active transportmechanisms mediated by efflux and uptake transporters that are expressedon the MDCK-MDR1 cells, predominantly by the overexpressed human MDR1P-gp. The compounds are assigned to permeability/absorption classes bycomparison of the AB permeabilities with the AB permeabilities ofreference compounds with known in vitro permeability and oral absorptionin the human. Identical or similar permeabilities in both transportdirections indicate passive permeation, vectorial permeability points toadditional active transport mechanisms. Higher PEBA than PEAB indicatesthe involvement of active efflux mediated by MDR1 P-gp. Active transportis concentration-dependently saturable.

MDCK-MDR1 cells (1-2×10e5 cells/1 cm2 area) are seeded on filter inserts(Costar transwell polycarbonate or PET filters, 0.4 μm pore size) andcultured (DMEM) for 7 days. Subsequently, the MDR1 expression is boostedby culturing the cells with 5 mM sodium butyrate in full medium for 2days. Compounds are dissolved in appropriate solvent (like DMSO, 1-20 mMstock solutions). Stock solutions are diluted with HTP-4 buffer (128.13mM NaCl, 5.36 mM KCl, 1 mM MgSO₄, 1.8 mM CaCl₂), 4.17 mM NaHCO₃, 1.19 mMNa₂HPO₄×7H₂O, 0.41 mM NaH₂PO₄xH₂O, 15 mM HEPES, 20 mM glucose, 0.25%BSA, pH 7.4) to prepare the transport solutions (0.1-300 μM compound,final DMSO <=0.5%). The transport solution (TL) is applied to the apicalor basolateral donor side for measuring A-B or B-A permeability (3filter replicates), respectively. The receiver side contains the samebuffer as the donor side. Samples are collected at the start and end ofexperiment from the donor and at various time intervals for up to 2hours also from the receiver side for concentration measurement byHPLC-MS/MS or scintillation counting. Sampled receiver volumes arereplaced with fresh receiver solution. Efflux ratio is calculateddividing the Papp (b-a) values by the Papp (a-b) values.

TABLE 5 Papp (PEBA) and efflux of compounds of the present inventionPapp (a-b) mean Ex. [10-6 cm/s] efflux ratio   1 78 0.6   2 84 0.6  3786 0.5  17 85 0.6  25 100 0.8  21 94 0.6   3 97 0.7  69 25 1.3  70 202.1  71 23 1.9  53 15 1.8 82a 59 1.0  51 11 5.0 81a 60 1.4  46 60 1.481b 64 1.2 83a 34 1.4 84a 31 2.2 84b 23 2.2

For the skilled in the art it is evident from the experimental resultsshown above that compounds of the present invention are not only potentphosphodiesterase 2 inhibitors but also have good membrane permeabilityand low to moderate in vitro efflux.

Metabolic Stability

The metabolic stability of the compounds according to the invention hasbeen investigated as follows:

The metabolic degradation of the test compound was assayed at 37° C.with pooled liver microsomes from various species. The final incubationvolume of 100 μl per time point contains TRIS buffer pH 7.6 at roomtemperature (0.1 M), magnesium chloride (5 mM), microsomal protein (1mg/mL for human and dog, 0.5 mg/mL for other species) and the testcompound at a final concentration of 1 μM. Following a shortpreincubation period at 37° C., the reactions were initiated by additionof betanicotinamide adenine dinucleotide phosphate, reduced form (NADPH,1 mM), and terminated by transferring an aliquot into solvent afterdifferent time points. After centrifugation (10000 g, 5 min), an aliquotof the supernatant was assayed by LC₁₀ MS/MS for the amount of parentcompound. The half-life was determined by the slope of thesemi-logarithmic plot of the concentration-time profile.

TABLE 4 Stability of compounds of the present invention in human livermicrosomes. Half-life—t½ Ex. [min] human   1 >130   2 120  37 >130 17 >130  25 >130  21 >130   3 72  68 >130  69 53  70 63  71 >130 51 >130  53 120 88b >130 82a >130 81a >130  46 >130 81b >130 83a >13086a >130

For the skilled in the art it is evident from the experimental resultsshown above that compounds of the present invention are not only potentphosphodiesterase 2 inhibitors but also have good metabolic stability.

In view of their ability to inhibit the activity of phosphodiesterase 2activity and their advantaneouges pharmacokinetics properties thecompounds of general formula (I) according to the invention, or thephysiologically acceptable salts thereof, are suitable for the treatmentand/or preventative treatment of all those diseases or conditions whichcan be influenced by inhibition of PDE2 hyperactivity and/or cAMP and/orcGMP hypofunction. Therefore, compounds according to the invention,including the physiologically acceptable salts thereof, are particularlysuitable for the prevention or treatment of diseases, particularly (1)disorders comprising the symptom of cognitive deficiency; (2) organic,including symptomatic, mental disorders, dementia; (3) mentalretardation; (4) mood affective disorders; (5) neurotic, stress-relatedand somatoform disorders including anxiety disorders; (6) behaviouraland emotional disorders with onset usually occurring in childhood andadolescence, attention deficit hyperactivity syndrome (ADHD) includingAutism spectrum disorders; (7) disorders of psychological development,developmental disorders of scholastic skills; (8) schizophrenia andother psychotic disorders; (9) disorders of adult personality andbehaviour; (10) mental and behavioural disorders due to psychoactivesubstance use; (11) extrapyramidal and movement disorders; (12) episodicand paroxysmal disorders, epilepsy; (13) Systemic atrophies primarilyaffecting the central nervous system, ataxia; (14) Behavioural syndromesassociated with physiological disturbances and physical factors; (15)sexual dysfunction comprising excessive sexual drive; (16) factitiousdisorders; (17) obsessive-compulsive disorders; (18) depression; (19)neuropsychiatric symptoms (e.g. depressive symptoms in Alzheimer'sdisease); (20) mixed dementia; (21) cognitive impairment inschizoaffective disorder; (22) cognitive impairment in bipolar disorderand (23) cognitive impairment in major depressive disorder.

In addition, the compounds of the present invention can be used for thetreatment, amelioration and/or prevention of cognitive impairment beingrelated to perception, concentration, cognition, learning, attention ormemory.

In addition, the compounds of the present invention can be used for thetreatment amelioration and/or prevention of cognitive impairment beingrelated to age-associated learning and memory impairments,age-associated memory losses, vascular dementia, craniocerebral trauma,stroke, dementia occurring after strokes (post stroke dementia),post-traumatic dementia, general concentration impairments,concentration impairments in children with learning and memory problems,Alzheimer's disease, Lewy body dementia, dementia with degeneration ofthe frontal lobes, including Pick's syndrome, Parkinson's disease,progressive nuclear palsy, dementia with corticobasal degeneration,amyotropic lateral sclerosis (ALS), Huntington's disease, multiplesclerosis, thalamic degeneration, Creutzfeld-Jacob dementia, HIVdementia, schizophrenia with dementia or Korsakoff's psychosis. Inaddition, the compounds of the present invention can be used for thetreatment of Alzheimer's disease.

In addition compounds of the present invention can be used for thetreatment of pain disorders, including but not limited to inflammatory,neuropathic and osteoarthritic pain.

In addition, the compounds of the present invention can be used for thetreatment of sleep disorders, bipolar disorder, metabolic syndrome,obesity, diabetes mellitus, hyperglycemia, dyslipidemia, impairedglucose tolerance, or a disease of the testes, brain, small intestine,skeletal muscle, heart, lung, thymus or spleen.

Preferably the compounds according to the invention are suitable for thetreatment of Alzheimer's Disease and for the treatment schizophrenia.

More preferably the compounds according to the invention are suitablefor symptomatic treatment of Alzheimer's Disease and for the treatmentof cognitive impairment associated with schizophrenia.

In particular the compounds according to the invention are suitable forsymptomatic treatment of prodromal and mild-to-moderate Alzheimer'sDisease and for the treatment of cognitive impairment associated withschizophrenia and symptomatic treatment of cognitive impairmentassociated with schizophrenia.

In a further aspect of the present invention the present inventionrelates to methods for the treatment or prevention of above mentioneddiseases and conditions, which method comprises the administration of aneffective amount of a compound of general formula (I), or thepharmaceutically acceptable salts thereof, to a human being.

The dose range of the compounds of general formula (I) applicable perday is usually from 0.1 to 1000 mg, preferably from 1 to 500 mg by oralroute, in each case administered 1 to 4 times a day.

Each dosage unit may conveniently contain from 0.1 to 500 mg, preferably1 to 100 mg.

The actual pharmaceutically effective amount or therapeutic dosage willof course depend on factors known by those skilled in the art such asage and weight of the patient, route of administration and severity ofdisease. In any case the combination will be administered at dosages andin a manner which allows a pharmaceutically effective amount to bedelivered based upon patient's unique condition.

Suitable preparations for administering the compounds of formula I,including the pharmaceutically acceptable salts thereof, will beapparent to those with ordinary skill in the art and include for exampletablets, pills, capsules, suppositories, lozenges, troches, solutions,syrups, elixirs, sachets, injectables, inhalatives, powders, etc. Thecontent of the pharmaceutically active compound(s) should be in therange from 0.1 to 95 wt.-%, preferably 5.0 to 90 wt.-% of thecomposition as a whole.

Suitable tablets may be obtained, for example, by mixing one or morecompounds according to formula I with known excipients, for exampleinert diluents, carriers, disintegrants, adjuvants, surfactants, bindersand/or lubricants. The tablets may also consist of several layers.

For this purpose, the compounds of formula I prepared according to theinvention may be formulated, optionally together with other activesubstances, together with one or more inert conventional carriers and/ordiluents, e.g. with corn starch, lactose, glucose, microcrystallinecellulose, magnesium stearate, citric acid, tartaric acid, water,polyvinylpyrrolidone, water/ethanol, water/glycerol, water/sorbitol,water/polyethylene glycol, propylene glycol, cetylstearyl alcohol,carboxymethylcellulose or fatty substances such as hard fat or suitablemixtures thereof.

The compounds according to the invention may also be used in conjunctionwith other active substances, particularly for the treatment and/orprevention of the diseases and conditions mentioned above. Other activesubstances which are suitable for such combinations include, forexample, BACE inhibitors; amyloid aggregation inhibitors (e.g.ELND-005); directly or indirectly acting neuroprotective and/ordisease-modifying substances; anti-oxidants (e.g. vitamin E orginkolide); anti-inflammatory substances (e.g. Cox inhibitors, NSAIDsadditionally or exclusively having Abeta lowering properties); HMG-CoAreductase inhibitors (statins); acetylcholinesterase inhibitors (e.g.,donepezil, rivastigmine, tacrine, galantamine); NMDA receptorantagonists (e.g. memantine); AMPA receptor agonists; AMPA receptorpositive modulators, AMPAkines, monoamine receptor reuptake inhibitors,substances modulating the concentration or release of neurotransmitters;substances inducing the secretion of growth hormone (e.g., ibutamorenmesylate and capromorelin); CB-1 receptor antagonists or inverseagonists; antibiotics (e.g., minocyclin or rifampicin); PDE2, PDE4,PDE5, PDE9, PDE10 inhibitors, GABAA receptor inverse agonists, GABAAreceptor antagonists, nicotinic receptor agonists or partial agonists orpositive modulators, alpha4beta2 nicotinic receptor agonists or partialagonists or positive modulators, alpha7 nicotinic receptor agonists orpartial agonists or positive modulators; histamine H3 antagonists, 5HT-4 agonists or partial agonists, 5HT-6 antagonists,alpha2-adrenoreceptor antagonists, calcium antagonists, muscarinicreceptor M1 agonists or partial agonists or positive modulators,muscarinic receptor M2 antagonists, muscarinic receptor M4 antagonists,metabotropic glutamate-receptor 5 positive modulators, glycinetransporter 1 inhibitors, antidepressants, such as citalopram,fluoxetine, paroxetine, sertraline and trazodone; anxiolytics, such aslorazepam and oxazepam; antiphychotics, such as aripiprazole, clozapine,haloperidol, olanzapine, quetiapine, risperidone and ziprasidone, andother substances that modulate receptors or enzymes in a manner suchthat the efficacy and/or safety of the compounds according to theinvention is increased and/or unwanted side effects are reduced.

The compounds according to the invention may also be used in combinationwith immunotherapies (e.g., active immunisation with Abeta or partsthereof or passive immunisation with humanised anti-Abeta antibodies ornanobodies) for the treatment of the above-mentioned diseases andconditions.

The dosage for the combination partners mentioned above is usefully 1/5of the lowest dose normally recommended up to 1/1 of the normallyrecommended dose.

Therefore, in another aspect, this invention relates to the use of acompound according to the invention or a pharmaceutically acceptablesalt thereof combined with at least one of the active substancesdescribed above as a combination partner, for preparing a pharmaceuticalcomposition which is suitable for the treatment or prevention ofdiseases or conditions which can be affected by inhibitors ofphosphodiesterase 2. These are preferably pathologies related to PDE2hyperactivity and/or cAMP and/or cGMP hypofunction, particularly one ofthe diseases or conditions listed above, most particularly prodromal andmild-to-moderate Alzheimer's Disease and cognitive impairment associatedwith schizophrenia. The use of the compound according to the inventionin combination with another active substance may take placesimultaneously or at staggered times, but particularly within a shortspace of time. If they are administered simultaneously, the two activesubstances are given to the patient together; while if they are used atstaggered times the two active substances are given to the patientwithin a period of less than or equal to 12 hours, but particularly lessthan or equal to 6 hours.

Consequently, in another aspect, this invention relates to apharmaceutical composition which comprises a compound according to theinvention or a pharmaceutically acceptable salt thereof and at least oneof the active substances described above as combination partners,optionally together with one or more inert carriers and/or diluents.

The compound according to the invention may both be present together inone formulation, for example a tablet or capsule, or separately in twoidentical or different formulations, for example as a so-calledkit-of-parts.

EXAMPLES

The following examples are intended to illustrate the invention, withoutrestricting its scope.

Chemical Manufacture

Abbreviations:

-   ACN acetonitrile-   APCI Atmospheric pressure chemical ionization-   d day-   Cy cyclohexane-   DCM dichloromethane-   DIPEA diisopropylethylamine-   DMF dimethylformamide-   ESI electrospray ionization (in MS)-   EtOAc ethylacetate-   EtOH ethanol-   Exp. Example-   GC gas chromathography-   GC-MS coupled gas chromatography-mass spectrometry-   h hour(s)-   HATU    O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-Hexafluorophosphate-   HCl hydrochloric acid-   HPLC high performance liquid chromatography-   HPLC-MS coupled high performance liquid chromatography-mass    spectrometry-   LC liquid chromatography-   LC-MS liquid chromatography—mass spectrometry-   M molar (mol/L)-   MeOH methanol-   min minute(s)-   MS mass spectrometry-   NaOH sodiumhydroxide-   NMP 1-methyl-2-pyrrolidinone-   NOE Nuclear Overhauser effect-   PE petroleum ether-   rt room temperature-   Rt retention time (in HPLC)-   HATU    1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium    3-oxid hexafluorophosphate-   TBTU O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium    tetrafluoroborate-   TEA triethylamine-   TFA trifluoroacetic acid-   THE tetrahydrofuran-   TLC thin-layer chromatography-   UPLC-MS ultra performance liquid chromatography—mass spectrometry

Analytical Methods:

UPLC-MS, HPLC-MS, LC-MS:

Method 1:

Instrument: LC/MS ThermoFinnigan HPLC Surveyor DAD, MSQ singlequadrupole

Column: Synergi Hydro RP100A, 2.5 μm, 3×50 mm

Mobile phase: A=H₂O 90%+10% CH₃CN+NH₄COOH 10 mM

-   -   B═CH₃CN 90%+H₂O 10%+NH₄COOH 10 mM

Time in min: % A % B Flow rate in mL/min 0.00 100 0 1.2 0.50 100 0 1.26.50 0 100 1.2 7.50 0 100 1.2 8.00 100 0 1.2 9.00 100 0 1.2

Detection: UV 254 nm

Detection: Finnigan MSQ, single quadrupole

Ion source: APCI+/APCI−

Scan range: 100-900 amu

Method 2:

Instrument: LC/MS Waters Acquity UPLC System DAD, SQD single quadrupole

Column: BEH C18 1.7 μm 2.1×50 mm, Temp 35° C.

Mobile phase: A=H₂O 90%+CH₃CN 10%+NH₄COOH 5 mM

-   -   B═CH₃CN 90%+H₂O 10%

Time in min: % A % B Flow rate in mL/min 0.00 100 0 0.7 1.20 0 100 0.71.45 0 100 0.7 1.55 100 0 0.7 1.75 100 0 0.7

Detection: UV 254 nm

Detection: SQD, single quadrupole

Ion source: ES+/ES−

Scan range: 90-900 amu

Method 3:

Instrument: LC/MS Waters Alliance 2695 HPLC System DAD, Quattro MicroTriple

quadrupole

Column: Atlantis dC18 5 m 4,6×50 mm, Temp 35° C.

Mobile phase: A=H₂O 90%+10% CH₃CN+CF₃COOH 0,05%

-   -   B═CH₃CN 90%+10% H₂O

Time in min: % A % B Flow rate in mL/min 0.00 100 0 1.3 0.70 100 0 1.34.50 0 100 1.3 5.80 0 100 1.3 6.00 100 0 1.3

Detection: UV 254 nm

Detection: Quattro Micro, triple quadrupole

Ion source: ES+

Scan range: 90-1000 amu

Method 4:

Instrument: LC/MS Waters Alliance 2695 HPLC System DAD, Quattro MicroTriple

quadrupole

Column: XBridge Phenyl 3.5 m 3×30 mm, Temp 35° C.

Mobile phase: A=H₂O 90%+10% CH₃CN+NH₄HCO₃ 5 mM

-   -   B═CH₃CN 90%+10% H₂O

Time in min: % A % B Flow rate in mL/min 0.00 100 0 1.3 4.50 0 100 1.35.80 0 100 1.3 6.00 100 0 1.3

Detection: UV 254 nm

Detection: Quattro Micro, triple quadrupole

Ion source: ES+

Scan range: 90-1000 amu

Method 5:

Instrument: LC/MS Waters Acquity UPLC System DAD, SQD single quadrupole

Column: BEH C18 1.7 μm 2.1×50 mm, Temp 35° C.

Mobile phase: A=H₂O 90%+CH₃CN 10%+NH₄HCO₃ 5 mM

-   -   B═CH₃CN 90%+H₂O 10%

Time in min: % A % B Flow rate in mL/min 0.00 100 0 0.70 1.20 100 0 0.701.45 0 100 0.70 1.55 0 100 0.70 1.75 100 0 0.70

Detection: UV 254 nm

Detection: SQD, single quadrupole

Ion source: ES+/ES−

Scan range: 90-900 amu

Method 6:

Instrument: LC/MS Waters Acquity System DAD, SQD single quadrupole

Column: XBridge C18 2.5 μm 3.0×30 mm, Temp 60° C.

Mobile phase: A=H₂O+TFA 0.1%

-   -   B═CH₃CN

Time in min: % A % B Flow rate in mL/min 0.00 98 2 1.5 1.3 1 99 1.5 1.51 99 1.5 1.6 98 2 1.5

Method 7:

Instrument: LC/MS Waters Acquity System DAD, SQD single quadrupole

Column: XBridge C18 2.5 μm 3.0×30 mm, Temp 60° C.

Mobile phase: A=H₂O+NH₄OH 0.1%

-   -   B═CH₃CN

Time in min: % A % B Flow rate in mL/min 0.00 95 5 1.5 1.3 1 99 1.5 1.51 99 1.5 1.6 95 5 1.5

Method 8:

Instrument: LC/MS Agilent 1100 System DAD

Column: Sunfire C18 2.5 μm 3.0×30 mm, Temp 60° C.

Mobile phase: A=H₂O+TFA 0.1%

-   -   B═CH₃CN

Time in min: % A % B Flow rate in mL/min 0.00 98 2.0 2.0 1.2 0.0 100 2.01.4 0.0 100 2.0

Method 10:

Instrument: LC/MS ThermoFinnigan HPLC Surveyor DAD, LCQFleet Ion Trap

Column: Xselect CSH, 2.5 μm, 4,6×50 mm

Mobile phase: A=H₂O 90%+10% CH₃CN+HCOOH 0.1%

-   -   B═CH₃CN 90%+H₂O 10%+HCOOH 0.1%

Time in min: % A % B Flow rate in mL/min 0.00 100 0 1.4 4.00 0 100 1.45.30 0 100 1.4 5.50 100 0 1.4 6.00 100 0 1.4

Detection: UV 254 nm

Detection: Finnigan Fleet, Ion Trap

Ion source: ES+

Scan range: 100-900 amu

GC/MS Method

Method 9:

Instrument: GC/MS Thermo Scientific TRACE GC ULTRA, DSQ II MS single

quadrupole

Column: Agilent DB-5MS, 25 m×0.25 mm×0.25 um

Carrier gas: Helium, 1 mL/min costant flow

Oven Program: 50° C., to 100° C. in 10° C./min, to 200° C. in 20°C./min, to 320° C. in

-   -   30° C./min (hold 10 min).

Detection: DSQ II MS single quadrupole

-   -   Ion source: El    -   Scan range: 50-450 amu

Chiral HPLC Methods:

Instrument: HPLC Agilent 1100 (DAD equipped; UV Detection: 230 nm); flowrate: 1 mL/min; column temperature: 25° C.

Method C1

column: Daicel Chiralpack AD-H; eluent: Hexane:lsopropanol=70:30

Method C2

column: Daicel Chiralpack AD-H; eluent: Hexane:lsopropanol=60:40

Method C3

column: Daicel Chiralpack AD-H; eluent: Hexane:lsopropanol=80:20

Method C4

column: Daicel Chiralcel OJ-H; eluent: Hexane:EtOH=80:20

Method C5

column: Daicel Chiralcel OJ-H; eluent: Hexane:EtOH=85:15

Method C6

column: Daicel Chiralcel OJ-H; eluent: Hexane:EtOH=70:30

Method C7

column: Daicel Chiralcel AS-H; eluent: Hexane:EtOH=75:25

NMR equipment:

The 1H NMR spectra were recorded on a Bruker Avance III (500 MHz) or aVarian 400 (400 MHz) or Varian Mercury (300 MHz) instrument usingdeuterated dimethylsulfoxide (DMSO-d6) as the solvent withtetramethylsilane (TMS) and residual solvent peak as an internalstandard. Chemical shifts are reported in δ values (ppm) relative toTMS.

Purification:

The most suitable purification techniques applied for the purificationof compounds of the present invention are direct phase silica gel flashchromatography and reverse phase chromatography, unless otherwisespecifically stated.

General Comment Concerning the Presentation of the Structures

Compounds with stereogenic centre(s): The structures depicted in theexperimental section will not necessarily show all the stereochemicalpossibilities of the compounds but only one.

The structural presentation of the compounds in the experimental sectionwill show a stereochemical bond only in case where the absolutestereochemistry is known.

The structural presentation of the compounds in the experimental sectionwith unknown absolute stereochemistry will show a planar bond plus anadditional comment that indicates if the described compound is a racemicmixture, a single stereoisomer and where applicable the relativestereochemistry.

Two examples are given below.

Example 1: The Presented Chemical Structure is Depicted as

Racemic Mixture

The added term racemic mixture points to the two stereochemical optionsand thus the manufactured compounds is a mixture of:

When racemic mixtures of above depicted structures are separated, thesingle stereoisomers are depicted as:

The added term ‘single stereoisomer’ and the planar bond indicates thatthe absolute configuration is unknown.

Single stereoisomer a is assigned to the first eluting isomer in chiralHPLC, single stereoisomer b is assigned to the second eluting isomer inchiral HPLC.

Example 2: The Presented Chemical Structure is Depicted as

The added term ‘TRANS-racemic mixture’ points to the two stereochemicaloptions and thus the manufactured compounds is a mixture of:

The same principles applies to ‘CIS-racemic mixture’.

When racemic mixtures of above depicted structures are separated, thesingle stereoisomers are depicted as:

The added term ‘TRANS-single stereoisomer’ indicates a relativeconfiguration known (trans) and the planar bond indicates the unknownabsolute configuration. The same principles applies to ‘CIS-singlestereoisomer’.

Single stereoisomer a is assigned to the first eluting isomer in chiralHPLC, single stereoisomer b is assigned to the second eluting isomer inchiral HPLC.

EXPERIMENTAL

The following intermediates and examples are intended to illustrate theinvention, without restricting its scope.

Intermediates

Intermediate 1:

To a solution of 3-amino-4-carbethoxypyrazole (4 g, 25.27 mmol) inabsolute EtOH (40 mL), 1,1,3,3-Tetraethoxy-2-methyl-propane (6.34 g,26.53 mmol) was added followed by 13.90 mL of a 1M solution of HCl indioxane. The mixture was heated at 80C overnight. Solvents wereevaporated, then DCM and water were added. Phases were separated,organics washed with a saturated solution of NaCl, dried over sodiumsulphate and evaporated to obtain 5.17 g of the title compound LC-MS(Method 2): R_(t)=0.73 min MS (ESI pos): m/z=206 (M+H)⁺

Intermediate 2

Intermediate 1 (5 g) was dissolved in a mixture of THE/water (1:1, 100mL) and stirred at room temperature for 48 hrs. The resulting suspensionwas diluted with water and 70 mL of EtOAc were added. Phases wereseparated, aqueous phases were treated with a 4N solution of HCl (ca 20mL). A white solid formed. The mixture was cooled at 0° C., then thewhite solid formed collected by filtration and dried under vacuum at 65°C. to obtain 3.50 g of the title compound.

LC-MS (Method 3): R_(t)=1.62 min

MS (ESI pos): m/z=178 (M+H)⁺

Intermediate 3:

To a solution of 2-Bromo-malonaldehyde (9.73 g; 64 mmol) in EtOH (100mL) at 70° C., 3-amino-4-carbethoxypyrazole (10 g, 64 mmol) and AcOH(100 mL) were added and the mixture stirred at 70° C. for 1 h. Solventswere evaporated, the residue treated with DCM (100 mL) and a 1 Nsolution of NaOH (100 mL). Phases were separated, organics washed with asaturated solution of NaCl, dried over sodium sulphate and evaporated.The crude was purified flash cromatography (eluent 10:1 PE/EtOAc) toobtain 15 g of the title compound as white solid.

LC-MS (Method 2): R_(t)=0.98 min MS (ESI pos): m/z=271 (M+H)⁺

Intermediate 4:

Intermediate 3 (5 g, 18.5 mmol) was suspended in dry toluene (50 mL) and5 mL of water were added. To this mixture, potassiumcyclopropyltrifluoroborate (4 g, 28 mmol) was added followed by2-dicyclohexylphosphino-2′,6′-di isopropoxy 1,1′-biphenyl (0.864 g, 1.85mmol), palladium acetate (0.208 g, 0.93 mmol) and potassium carbonate(7.7 g, 55 mmol). Mixture was refluxed at 130° C. for 3 hrs, then cooledto room temperature, filtered over celite and washed with AcOEt and thenEtOH. Solvent was evaporated under vacuum and the crude used in the nextstep without further purification.

LC-MS (Method 2): R_(t)=0.9 min

MS (ESI pos): m/z=232 (M+H)⁺

Intermediate 5:

Intermediate 4 (4 g, 17.5 mmol) was suspended in 50 ml of EtOH, 8 ml of4N NaOH and 30 ml of water and stirred overnight. EtOH was evaporatedand a 4 N solution of HCl added. The solid formed was filtered, washedwith water and dried under vacuum at 70° C. overnight to obtain 3.6 g ofthe title compound.

LC-MS (Method 3): R_(t)=2.75 min

MS (ESI pos): m/z=204 (M+H)⁺

Intermediate 6:

Intermediate 6 was prepared as described in WO 2010/007074 starting fromcommercially available (Z) 3-(diethylamino)-2-fluoroprop-2-enal (1.34mL, 9.0 mmol) and 3-amino-4-carbethoxypyrazole (2.1 g, 13.6 mmol) toobtain 0.53 g of the title compound.

¹H NMR (300 MHz, CDCl3): δ ppm 1.44-1.39 (t, 3H), 4.47-4.40 (q 2H) 8.57(s, 1H) 8.7 (m, 1H), 8.8 (d, 1H) Intermediate 7:

To a solution of 5-amino-3-methyl-1H-pirazole-4-carboxylic acid ethylester (1 g, 5.91 mmol) in absolute EtOH (25 mL),1,1,3,3-Tetraethoxy-2-methyl-propane (1.4 g, 6.2 mmol) was addedfollowed by 1.63 mL of a 4N solution of HCl in dioxane.Mixture washeated at 80° C. for 5 hrs, left at room temperature overnight and thensolvents were evaporated to dryness. The violet solid obtained wasdissolved in DCM, water was added and the phases separated.

The organic phases were dried over sodium sulfate and concentrated undervacuum to obtain 1.26 g of title compound used for next step withoutfurther purification.

LC-MS (Method 2): R_(t)=0.79 min

MS (ESI pos): m/z=224 (M+H)⁺

Intermediate 8:

To a solution of intermediate 7 (1.26 g, 5.75 mmol) in THE (25 mL) andwater (25 mL) 1.5 mL of a 1 N solution of sodium hydroxide were addedand the mixture heated at heated at 60° C. for 2 h. Solvent wasevaporated, water was added and 30 ml of a 12N solution of HCl addeduntil pH 2. The solid formed was filtered, washed with water and driedat 70° C. under vacuum to obtain 0.9 g of title compound as white solid.

LC-MS (Method 1): R_(t)=0.27 min

MS (APCI): m/z=192 (M+H)⁺

Intermediate 9:

To a solution of 5-amino-3-methyl-1H-pyrazole-4-carboxilic acid ethylester (4 g, 23.64 mmol) in absolute EtOH (80 mL),1,1,3,3-tetraethoxypropane (5.96 mL, 23.64 mmol) and 5.9 mL of a 4Nsolution of HCl in dioxane were added. The resulting mixture was heatedat 80° C. 3 hrs. Solvents were evaporated, the residue was diluted withDCM and water. Organic layer was separated, dried over sodium sulphateand evaporated to obtain the title compound as white solid (3.6 g)

LC-MS (Method1): R_(t)=263 min

MS (APCI): m/z=206 (M+H)⁺

Intermediate 10:

To a solution of intermediate 9 (3.6 g, 17.54 mmol) in AcOH (70 mL)bromine (2.26 mL) was added drowpwise. The mixture was stirred at roomtemperature overnight then carefully poured into 500 mL of water andextracted with EtOAc (3×100 mL). Organic phases were collected andwashed with 100 mL of a 5% solution of Na2S2O3 and then with 100 mL of asaturated solution of NaCl, dried over sodiumsulphate and concentratedunder vacuum.

Crude was purified by flash chromatography (eluent: DCM/EtOAc; gradientfrom 100% to 70%) to obtain the title compound as white solid (2.1 g)LC-MS (Method 1): R_(t)=3.52 min MS (APCI): m/z=284 (M+H)⁺

Intermediate 11:

To a solution of intermediate 10 (2.05 g, 7.22 mmol) in toluene (40 mL)water (4 mL) was added followed by potassium cyclopropyltrifluroborate(1.6 g, 10.82 mmol), palladium(II) acetate (0.08 g, 0.36 mmol),dicyclohexylphosphino-2′,6′-di-i-propoxy dl-1,1′-biphenyl (RUPHOS, 0.34g, 0.72 mmol) and potassium carbonate (3 g, 21.65 mmol). Mixture washeated at 130° C. for 3 hrs then cooled to room temperature, filteredover celite and washed with AcOEt. Organic layer was dried andevaporated to obtain the title compound (1.5 g) used for the next stepwithout further purification.

LC-MS (Method 2): R_(t)=0.92 min

MS (ESI pos): m/z=246 (M+H)⁺

Intermediate 12:

To a solution of intermediate 11 (1.5 g, 6.2 mmol) in absolute EtOH (30mL) water (10 mL) was added followed by 7.7 mL of a 8N solution of NaOH.Mixture was stirred at room temperature overnight, then solventevaporated and a 4N solution of HCl added until pH=1. The solid formedwas filtered, washed with water and dried under vacuum at 70° C.overnight (1.5 g).

LC-MS (Method 1): R_(t)=0.6 min

MS (APCI): m/z=218 (M+H)⁺

Intermediate 13:

To a solution of 4-bromo-3-fluoro-benzotrifluoride (585 mg, 2.36 mmol)in 15 mL of THF, stirred at −75° C. under nitrogen atmosphere, 1.53 mL(2.6 mmol) of a 1.7M solution of tert-butyllithium in pentane were addeddropwise. The reaction mixture was stirred at −60° C. for 15 minutesthen a solution of 2-methyl-propane-2-sulfinicacid-(tetrahydro-pyran-4-ylidene)-amide(400 mg, 1.97 mmol; prepared asdescribed in literature: WO2005/87751 A2) in 10 mL of THE was addeddropwise. The reaction mixture was allowed to reach room temperature andstirred for 1 hr. A saturated ammonium chloride solution was added andthe reaction mixtures was extracted with ethyl acetate. The organicphases were collected, dried over sodium sulfate and concentrated undervacuum. The crude obtained was purified by flash chromatography (eluent:cyclohexane/AcOEt; gradient from 12% to 100% of AcOEt). The oil obtainedwas diluted in 2 mL of 1,4-dioxane, 0.4 mL of a 4 M solution ofhydrochloric acid in 1,4-dioxane were added, the reaction mixture wasstirred at room temperature for 1 hr and then concentrated under vacuumto obtain 100 mg of the title compound as white solid.

LC-MS (Method 2): R_(t)=0.90 min

MS (ESI pos): m/z=264 (M+H)⁺

The following Amine Intermediates were prepared in analogy toIntermediate 13 starting from the corresponding commercially availablebromo-aryl/heteroaryl or iodo-aryl/heteroaryl derivative:

R_(t) Starting Amine intermediate MS m/z (min) Method 4-Chloro-2-fluoro- iodobenzene 14

230, 232 (M + H)+ 0.79 Method 2 4-Iodo- benzo- trifluoride 15

246 (M + H)+ 0.66 Method 2 — 16 Commercially available from ENAMINE-BB(Cat. Number EN300- 185595)

— — — 2-Bromo- quinoline 17

229 (M + H)+ 0.69 Method 2 — 18 Commercially available from ENAMINE-BB(Cat. Number EN300- 50665)

— — — 4-Bromo- 1- (difluoro- methoxy)-2- fluorobenz- ene 19

262 (M + H)+ 0.68 Method 2 4-Chloro- 3-fluoro- iodobenzene 20

230, 232 (M + H)+ 0.77 Method 2 2-Bromo- 3-fluoro-5- (trifluorometh-yl)pyridine 21

265 (M + H)+ 0.83 Method 2 2-Iodo-5- (trifluoro- methyl)pyridine 22

247 (M + H)+ 0.75 Method 2 5-Iodo-2- (trifluoro- methyl)pyridine 23

247 (M + H)+ 0.71 Method 2 5-Bromo- 3-fluoro-2- (trifluoro-methyl)pyridine 24

265 (M + H)+ 0.81 Method 2 2-Chloro- 4-fluoro- iodobenzene 25

230, 232 (M + H)+ 0.59 Method 2

Intermediate 26:

Intermediate 26 was prepared as described for Intermediate 13 startingfrom commercially available 4-bromo-3-fluoro-benzotrifluoride (560 mg,2.30 mmol) and 2-methyl-propane-2-sulfinic acid[dihydro-pyran-(3Z)-ylidene]-amide (390 mg, 1.92 mmol; prepared inanalogy to 2-methyl-propane-2-sulfinicacid-(tetrahydro-pyran-4-ylidene)-amide, described in WO2005/87751 A2)to obtain 120 mg of the title compound, as racemic mixture.

LC-MS (Method 2): R_(t)=1.00 min

MS (ESI pos): m/z=264 (M+H)⁺

The following Amine Intermediates were prepared in analogy toIntermediate 26 starting from the corresponding commercially availablebromo-aryl derivative:

R_(t) Starting Amine intermediate MS m/z (min) Method 4-Bromo- benzotri-fluororide 27 Racemic mixture

246 (M + H)+ 0.92 Method 2 1-Bromo- 4- (trifluoro- methoxy) benzene 28Racemic mixture

262 (M + H)+ 0.95 Method 2

Intermediate 29:

Intermediate 29 was prepared as described for Intermediate 13 startingfrom commercially available 4-bromo-3-fluoro-benzotrifluoride (462 g,1.90 mmol) and 2-methyl-propane-2-sulfinic acid[dihydro-furan-(3Z)-ylidene]-amide (300 mg, 1.58 mmol; prepared inanalogy to 2-methyl-propane-2-sulfinicacid-(tetrahydro-pyran-4-ylidene)-amide, described in WO2005/87751 A2)to obtain 50 mg of the title compound, as racemic mixture.

LC-MS (Method 2): R_(t)=0.90 min

MS (ESI pos): m/z=250 (M+H)⁺

The following Amine Intermediates were prepared in analogy toIntermediate 29 starting from the corresponding commercially availablebromo-aryl/heteroaryl or iodo-aryl/heteroaryl derivative:

R_(t) Starting Amine intermediate MS m/z (min) Method 4- Iodobenzo-trifluoride 30 Racemic mixture

232 (M + H)+ 0.83 Method 2 4-Chloro- 2- fluoroiodo- benzene 31 Racemicmixture

216, 218 (M + H)+ 0.75 Method 2 2-Bromo- 3-fluoro-5- (trifluoro-methyl)pyridine 32 Racemic mixture

251 (M + H)+ 0.76 Method 2 5-Iodo-2- (trifluoro- methyl)pyridine 33Racemic mixture

233 (M + H)+ 0.68 Method 2 2-Iodo-5- (trifluoro- methyl)pyridine 34Racemic mixture

233 (M + H)+ 0.70 Method 2

Intermediate 35:

Step 1:

3.6-Dihydro-2H-pyran-4-boronic acid pinacol ester (5.62 g, 26.75 mmol),4-bromo-3-fluorobenzotrifluoride (5.00 g, 20.58 mmol), potassiumcarbonate (8.53 g, 61 0.73 mmol) and1,1′-bis(diphenylphosphino)ferrocenedichloro palladium(II) (753 mg, 10.03 mmol) were suspended in 50 mL of 1,4-dioxane and 10 mL of water.The reaction mixture was refluxed for 3 hrs, solvents were evaporatedand the crude was extracted with ethyl acetate (50 mL) and water (50mL). Organic layer was separated, dried over sodium sulfate andevaporated. The crude obtained was purified by flash chromatography(eluent: cyclohexane/AcOEt; gradient from 40% to 100% of AcOEt) toobtain 4.0 g of the title compound as clear oil.

GC-MS (Method 9): R_(t)=7.76 min

MS: m/z=246 (M)⁺

Step 2:

To a solution of4-(2-Fluoro-4-trifluoromethyl-phenyl)-3,6-dihydro-2H-pyran (obtained asdescribed in Step 1; 7.0 g, 25.18 mmol) in 150 mL of dichloromethane,stirred at 0° C., 3-chloroperoxybenzoic acid (11.3 g, 50.37 mmol) wasadded portionwise. The reaction mixture was allowed to reach roomtemperature and stirred overnight. The reaction mixture was cooled to 0°C. and the precipitate formed was filtered off. The organic solution waswashed twice with an aqueous saturated solution of potassium carbonate,dried over sodium sulfate and concentrated under vacuum. The crudeobtained was purified by flash chromatography (eluent:cyclohexane/AcOEt; gradient from 50% to 100% of AcOEt) to obtain 4.2 gof the title compound.

GC-MS (Method 9): R_(t)=7.68 min

MS: m/z=262 (M)⁺

Step 3:

To a solution of6-(2-Fluoro-4-trifluoromethyl-phenyl)-3,7-dioxa-bicyclo[4.1.0]heptane(obtained as described in Step 2; 1.64 g, 6.25 mmol) in 10 mL ofacetonitrile, stirred under nitrogen atmosphere at −45° C.,trifluoromethane sulfonic acid (1.88 g, 12.5 mmol) was added dropwise.The reaction mixture was allowed to reach room temperature and stirredfor 2.5 hrs. 10 mL of water were added, the reaction mixture was warmedto 100° C. and acetonitrile was distilled out. The reaction mixture wasstirred at 100° C. for 5 hrs, then cooled to room temperature andstirred overnight. The reaction mixture was diluted with dichloromethaneand phases were separated. The aqueous phase was treated with a 4Msolution of NaOH until basic pH and extracted with dichloromethane. Theorganic phase was dried over sodium sulfate and concentrated undervacuum to give 290 mg of the final compound (crude colorless oil),4-Amino-4-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-pyran-3-ol, asracemic mixture (TRANS/CIS diastereoisomeric ratio 85:15, determined byNMR).

The crude was used in the next step without any further purification.

LC-MS (METHOD 2): R_(t)=0.77 min

MS (ESI pos): m/z=280 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b 7.59-7.47 (m, 3H), 4.74 (d, J=5.8 Hz, 1H),4.04 (dd, J=11.7, 1.5 Hz, 1H), 3.90 (ddd, J=12.5, 11.0, 2.0 Hz, 1H),3.74 (d, J=5.8 Hz, 1H), 3.67 (dd, J=11.1, 4.1 Hz, 1H), 3.56-3.51 (m,1H), 2.51-2.44 (m, 1H), 2.09 (br s, 1H), 1.56 (m, 1H).

NOE: 2.09 (NH2): 3.74; 4.04. 4.74 (OH): 3.55; 2.45

The following Amino-alcohol Intermediate were prepared in analogy toIntermediate 34 starting from the corresponding commercially availablebromo-heteroaryl:

R_(t) Starting Amino-alcohol intermediate MS m/z (min) Method 5-Bromo-2- (trifluoro- methyl)pyridine 36 Trans racemate

263 (M + H)+ 0.90 Method 1

Relative stereochemistry of intermediate 36 assigned by NMR and NOE:

1H NMR (500 MHz, DMSO-d6) b 8.87 (d, J=2.3 Hz, 1H, 13), 8.10 (ddd,J=8.3, 2.4, 0.8 Hz, 1H), 7.81 (dd, J=8.3, 0.8 Hz, 1H), 4.78 (d, J=5.9Hz, 1H), 4.05 (dd, J=11.6, 1.5 Hz, 1H), 3.91 (td, J=11.7, 2.3 Hz, 1H),3.68 (ddd, J=11.1, 4.9, 2.2 Hz, 1H), 3.56 (dd, J=11.7, 2.4 Hz, 1H), 3.47(d, J=5.9 Hz, 1H), 2.49-2.44 (m, 1H), 2.12 (s, 2H), 1.49 (dd, J=13.1,1.9 Hz, 1H).

NOE:2.12 (NH2): 3.47; 3.91;4.05. 4.78 (OH): 3.56; 2.48

Intermediate 37

Step 1:

tert-Butyllithium (21.8 mL, 1.7M in pentane, 37.0 mmol) was addeddropwise to 4-bromo-3-fluorobenzotrifluoride (5.00 g, 20.58 mmol) in THE(50 mL) at −70° C. After 1 h, 3-oxotetrahydrofuran (1.78 g, 20.58 mmol)in THE was added dropwise. The reaction mixture was warmed to −10° C.mixture and quenched with NH₄Cl satured solution. Ethyl acetate wasadded, the organic layer was separated, dried over sodium sulfate andevaporated. The crude obtained was purified by flash chromatography(eluent: cyclohexane/AcOEt; gradient from 0% to 80% of AcOEt) to obtain1.6 g of the 3-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-furan-ol.

GC-MS (Method 9): R_(t)=7.58 min

MS: m/z=250 (M)⁺

Step 2:

p-Toluenesulfonic acid monohydrate (1.75 g, 9.19 mmol) was added to3-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-furan-ol (obtained asdescribed in Step 1; 2.3 g, 9.19 mmol) in toulene (20 mL). Afterrefluxing for 1 h, volatiles were evaporated, DCM and water were added,the organic layer was separated, dried over sodium sulfate andevaporated to obtain 2.0 g (77% content) of crude3-(2-fluoro-4-trifluoromethyl-phenyl)-2,5-dihydro-furan, that was usedwithout further purification.

GC-MS (Method 9): R_(t)=7.12-7.21 min

MS: m/z=232 (M)⁺

Step 3:

To a solution of 3-(2-fluoro-4-trifluoromethyl-phenyl)-2,5-dihydro-furan(obtained as described in Step 2; 2.0 g 77% content, 6.63 mmol) in 50 mLof dichloromethane, stirred at 0° C., 3-chloroperoxybenzoic acid (2.63g, 15.26 mmol) was added portionwise. The reaction mixture was allowedto reach room temperature and stirred overnight. The reaction mixturewas cooled to 0° C. and the precipitate formed was filtered off. Theorganic solution was washed twice with an aqueous saturated solution ofpotassium carbonate, dried over sodium sulfate and concentrated undervacuum. The crude obtained was purified by flash chromatography (eluent:cyclohexane/AcOEt; gradient from 50% to 100% of AcOEt) to obtain 1.2 g(98% content) of1-(2-fluoro-4-trifluoromethyl-phenyl)-3,6-dioxa-bicyclo[3.1.0]hexane. ¹HNMR (300 MHz, DMSO-d6): b ppm 3.91-3.95 (m, 2H), 4.06-4.19 (m, 3H) 7.37(dd, J=10.2, 1.3 Hz,1H), b 7.47 (dd, J=8.4, 1.1 Hz, 1H), 7.59 (m, 1H)

Step 4:

To a solution of1-(2-fluoro-4-trifluoromethyl-phenyl)-3,6-dioxa-bicyclo[3.1.0]hexane(obtained as described in Step 3; 1.20 g, 98% content, 4.74 mmol) in 20mL of acetonitrile, stirred under nitrogen atmosphere at −40° C.,trifluoromethane sulfonic acid (0.84 mL, 9.48 mmol) was added dropwise.The reaction mixture was allowed to reach room temperature and stirredfor 2.5 hrs. 20 mL of water were added, the reaction mixture was warmedto 100° C. and acetonitrile was distilled out. The reaction mixture wasstirred at 100° C. for 20 hrs, then cooled to room temperature andstirred overnight. The reaction mixture was diluted with dichloromethaneand phases were separated. The aqueous phase was treated with a 4Msolution of NaOH until basic pH and extracted with dichloromethane. Theorganic phase was dried over sodium sulfate and concentrated undervacuum to give 200 mg of4-amino-4-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-furan-3--ol, asracemic mixture (TRANS/CIS diastereoisomeric ratio 88/12, determined byNMR).

The crude was used in the next step without any further purification.

LC-MS (METHOD 1): R_(t)=2.52-3.04 min

MS (ESI pos): m/z=266 (M+H)⁺

1H NMR (400 MHz, DMSO-d6) b 7.59-7.54 (m, 1H), 7.52-7.45 (m, 2H), 5.08(d, J=4.8 Hz, 1H), 4.29 (q, J=3.8 Hz, 1H), 4.25 (dd, J=8.8, 3.8 Hz, 1H),4.14 (d, J=7.6 Hz, 1H), 3.95 (dd, J=8.0, 2.2 Hz, 1H), 3.65 (d, J=8.8 Hz,1H), 2.06 (s, 2H).

NOE: 2.06 (NH2): 3.95; 4.29; 4.25. 5.08 (OH): 4.14; 3.65

Intermediate 38:

Step 1:

6-(2-Fluoro-4-trifluoromethyl-phenyl)-3,7-dioxa-bicyclo[4.1.0]heptane(obtained as described in Step 2 in the preparation of Intermediate 34;4.20 g, 16.02 mmol), sodium azide (2.08 g, 32.04 mmol) and ammoniumchloride (1.72 g, 32.04 mmol) were suspended in 50 mL of methanol and 10mL of water. The reaction mixture was stirred at reflux for 18 hrs.Solvents were removed, the crude was suspended in water and extractedtwice with ethyl acetate. The organic layer was dried over sodiumsulfate and concentrated under vacuum to give 4.70 g of the finalcompound as a mixture of the desired regioisomer4-Azido-4-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-pyran-3-ol andthe undesired regioisomer3-azido-4-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-pyran-4-ol in aregioisomeric ratio of 76/24 determined by NMR. The regioisomericmixture was used in the next step without separation.

GC-MS (METHOD 9): R_(t)=9.57 min

MS: m/z=248 (M)⁺

1H NMR (500 MHz, DMSO-d6) b 7.77-7.74 (m, 1H), 7.72 (d, J=7.9 Hz, 1H),7.67-7.64 (m, 1H), 5.37 (d, J=6.0 Hz, 1H), 3.93-3.89 (m, 1H), 3.85-3.80(m, 1H), 3.78 (dd, J=12.3, 1.5 Hz, 1H), 3.74-3.66 (m, 2H), 2.65 (ddd,J=13.8, 11.9, 4.8 Hz, 1H), 2.02-1.95 (m, 1H).

Step 2:

A mixture of4-Azido-4-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-pyran-3-ol and3-azido-4-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-pyran-4-ol(obtained as described in Step 1, 2.0 g, 6.55 mmol), Pd/C (300 mg, 2.82mmol) and di-tert-butyldicarbonate (1.86 g, 8.52 mmol) were suspended in150 mL of ethanol. The reaction mixture was stirred at room temperatureunder hydrogen atmosphere (2.5 bar) for 1 hr. The reaction mixture wasfiltered on a celite pad and the organic solution was concentrated undervacuum. The crude obtained was purified by flash chromatography (eluent:cyclohexane/AcOEt; gradient from 10% to 100% of AcOEt) to obtain 1.75 gof the title compound (yellow solid) as a mixture of the desiredregioisomer[4-(2-fluoro-4-trifluoromethyl-phenyl)-3-hydroxy-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester and the undesired regioisomer[4-(2-Fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-tetrahydro-pyran-3-yl]-carbamicacid tert-butyl ester in a regioisomeric ratio of 85/15 determined byNMR. The regioisomeric mixture was used in the next step withoutseparation.

GC-MS (METHOD 9): R_(t)=10.92-10.99 min

MS: m/z=323 (M)⁺

Step 3:

A mixture of[4-(2-Fluoro-4-trifluoromethyl-phenyl)-3-hydroxy-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester and4-(2-Fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-tetrahydro-pyran-3-yl]-carbamicacid tert-butyl ester (obtained as described in Step 2, 3.7 g, 7.30mmol) was dissolved in 20 mL of dichlorometane, Dess-Martin periodinane(2.18 g, 9.5 mmol) was added portionwise and the reaction mixture wasstirred at room temperature for 2 hrs. The reaction mixture was dilutedwith dichloromethane, washed with aqueous bicarbonate saturatedsolution, washed with aqueous sodium bisulfite saturated solution, theorganic layer was separated, dried over sodium sulfate and concentratedunder vacuum. The crude obtained was purified by flash chromatography(eluent: dichloromethane/AcOEt; gradient from 0% to 70% of AcOEt) togive 2.4 g of the desired compound.

LC-MS (METHOD 1): R_(t)=4.23-4.83 min

MS (ESI pos): m/z=278 (fragment) (M+H)⁺

Step 4:

[4-(2-Fluoro-4-trifluoromethyl-phenyl)-3-oxo-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester (obtained as described in Step 3; 340 mg, 0.9mmol) was suspended in 10 mL of dry THF. The reaction mixture wasstirred at −20° C. and 0.29 mL of a 3.4M solution of methylmagnesiumbromide in methyl-tetrahydrofurane was added dropwise. The reactionmixture was stirred at −20° C. for 1 hr, then quenched with aqueoussaturated ammonium chloride solution. Organic layer was separated, driedover sodium sulfate and concentrated under vacuum. The crude obtainedwas purified by flash chromatography (eluent: dichloromethane/AcOEt;gradient from 0% to 30% of AcOEt) to give 200 mg of the title compound,4-(2-fluoro-4-trifluoromethyl-phenyl)-3-hydroxy-3-methyl-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester, as racemic mixture (TRANS/CIS diastereoisomericratio 82/12, determined by NMR).

GC-MS (METHOD 9): R_(t)=11.01 min

MS: m/z=292 (fragment) (M)⁺

1H NMR (500 MHz, DMSO-d6) b 7.58-7.53 (m, 1H), 7.52-7.48 (m, 1H), 7.45(d, J=12.8 Hz, 1H), 6.92 (s, 1H), 4.78 (s, 1H), 3.88 (d, J=12.2 Hz, 1H),3.70 (t, J=6.7 Hz, 1H), 3.60 (q, J=12.8, 12.2 Hz, 1H), 3.28-3.26 (d,J=12.2 Hz, 1H), 2.88 (t, J=11.4 Hz, 1H), 1.33 (s, 9H), 0.9 (s, 3H).

NOE: 6.92 (NH): 0.90; 3.88. 4.78 (OH): 2.88; 3.27

Step 5:

4-(2-Fluoro-4-trifluoromethyl-phenyl)-3-hydroxy-3-methyl-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester (obtained as described in Step 4, as preferreddiastereoisomer; 200 mg, 0.51 mmol) was dissolved in 5 mL ofdichloromethane. Trifluoroacetic acid (0.39 mL, 5.1 mmol) was added, thereaction mixture was stirred at room temperature for 1 hr and thenconcentrated under vacuum. The crude obtained was stripped twice withethyl ether to give 198 mg of the title compound,4-amino-4-(2-fluoro-4-trifluoromethyl-phenyl)-3-methyl-tetrahydro-pyran-3-oltrifluoroacetate salt as racemic mixture (TRANS/CIS diastereoisomericratio 85/15, determined by NMR).

LC-MS (METHOD 1): R_(t)=3.35 min

MS (ESI pos): m/z=294 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b 8.61 (s, 3H), 7.96 (t, J=8.2 Hz, 1H), 7.81(dd, J=13.3, 2.0 Hz, 1H), 7.70-7.66 (m, 1H), 5.40 (s, 1H), 3.98 (ddd,J=13.3, 10.7, 2.8 Hz, 1H), 3.92-3.85 (m, 1H), 3.62 (d, J=12.6 Hz, 1H),3.40-3.38 (d, J=12.6 Hz, 1H), 2.99 (ddd, J=14.4, 10.7, 5.2 Hz, 1H), 1.79(dt, J=14.4, 3.0 Hz, 1H), 1.11 (d, J=1.8 Hz, 3H).

Intermediate 39:

Step 1:

Step 1 was performed in analogy to Step 1 in the preparation ofIntermediate 35, starting from2-bromo-3-fluoro-5-(trifluoromethyl)pyridine (5 g, 20.49 mmol) to obtain2-(3,6-dihydro-2H-pyran-4-yl)-3-fluoromethyl-pyridine (5.7 g).

LC-MS (METHOD 2): R_(t)=1.17 min

MS (ESI pos): m/z=248 (M+H)⁺

Step 2:

Step 2 was performed in analogy to Step 2 in the preparation ofIntermediate 35, starting from2-(3,6-Dihydro-2H-pyran-4-yl)-3-fluoromethyl-pyridine (5.7 g, 23.06mmol) to obtain2-(3,7-dioxa-bicyclo[4.1.0]hept-6-yl)-3-fluoro-5-trifluoromethyl-pyridine(3.25 g).

LC-MS (METHOD 2): R_(t)=0.95 min

MS (ESI pos): m/z=264 (M+H)⁺

Step 3:

Step 3 was performed in analogy to Step 1 in the preparation ofIntermediate 38, starting from2-(3,7-Dioxa-bicyclo[4.1.0]hept-6-yl)-3-fluoro-5-trifluoromethyl-pyridine(250 mg, 0.95 mmol) to obtain after purification by flash chromatography(eluent: cyclohexane/EtOAc; gradient from 0% to 30% of EtOAc),4-azido-4-(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-tetrahydro-pyran-3-ol(160 mg) as major regioisomer

LC-MS (METHOD 2): R_(t)=1.05 min

MS (ESI pos): m/z=307 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b 8.90 (dq, J=2.0, 0.9 Hz, 1H), 8.39 (ddd,J=11.5, 1.9, 0.7 Hz, 1H), 5.42 (s, 1H), 4.01 (s, 1H), 3.93-3.86 (m, 1H),3. 78-3.64 (m, 3H), 2.73 (ddd, J=14.6, 12.6, 4.9 Hz, 1H), 2.02 (dq,J=14.7, 2.0 Hz, 1H).

The minor regioisomer,3-Azido-4-(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-tetrahydro-pyran-4-ol,was also isolated (40 mg).

LC-MS (METHOD 2): R_(t)=1.04 min

MS (ESI pos): m/z=307 (M+H)⁺

Step 4:

To a solution of4-azido-4-(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-tetrahydro-pyran-3-ol(160 mg, 0.52 mmol) in 5 mL of methanol stirred under nitrogenatmosphere, ammonium formate (165 mg, 2.61 mmol) and zinc (51.2 mg, 0.78mmol) were added. The reaction mixture was stirred at room temperatureovernight and concentrated. A saturated ammonium chloride water solutionwas added and the reaction mixture was extracted with dichloromethane.The organic phase was separated, washed with brine, dried over sodiumsulfate and concentrated under vacuum to give 115 mg of4-Amino-4-(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-tetrahydro-pyran-3-ol,as TRANS-racemic mixture.

LC-MS (METHOD 5): R_(t)=0.71 min

MS (ESI pos): m/z=281 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) δ 8.75 (dq, J=2.0, 1.0 Hz, 1H), 8.18-8.12 (m,1H), 4.80 (d, J=5.6 Hz, 1H), 4.01 (dd, J=11.7, 1.5 Hz, 1H), 3.89-3.83(m, 1H), 3.78 (dt, J=5.5, 1.9 Hz, 1H), 3.74-3.66 (m, 1H), 3.55 (dd,J=11.7, 1.7 Hz, 1H), 2.71-2.61 (m, 1H), 2.09 (s, 2H), 1.65-1.58 (m, 1H).

NOE: 2.09 (NH2): 3.55; 3.70; 3.78 4.80 (OH): 1.61; 3.78

The following intermediate was prepared in analogy to Intermediate 39,starting from 2-Bromo-5-trifluoromethyl-pyridine

R_(t) Starting Amino-alcohol intermediate MS m/z (min) Method 2-Bromo-5-trifluoromethyl pyridine 40

263 0.68 METHOD 2

1H NMR (500 MHz, DMSO-d6) δ 8.89 (dq, J=2.6, 0.9 Hz, 1H), 8.13 (ddd,J=8.4, 2.5, 0.8 Hz, 1H), 7.69 (dt, J=8.4, 0.8 Hz, 1H), 4.69 (d, J=5.6Hz, 1H), 4.05 (dd, J=11.4, 1.7 Hz, 1H), 3.84 (td, J=11.3, 2.4 Hz, 1H),3.72 (ddd, J=10.9, 4.7, 2.7 Hz, 1H), 3.60 (ddd, J=5.7, 2.8, 1.4 Hz, 1H),3.54 (dd, J=11.5, 2.8 Hz, 1H), 2.56-2.51 (m, 1H), 2.02 (s, 2H),1.62-1.52 (m, 1H).

NOE: 2.09 (NH2): 3.55; 3.70; 3.78 4.80 (OH): 1.61; 3.78

Intermediate 41:

Step 1:

Step 1 was performed in analogy to Step 1 in the preparation ofIntermediate 38, starting from1-(2-fluoro-4-trifluoromethyl-phenyl)-3,6-dioxa-bicyclo[3.1.0]hexane(750, 3.02 mmol, prepared as described in Step 3 in the preparation ofIntermediate 37) to obtain4-azido-4-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-furan-3-ol asmajor regioisomer and4-Azido-3-(2-fluoro-4-trifluoromethyl-phenyl)-tetrahydro-furan-3-ol asminor regioisomer. (900 mg, regioisomer ratio 82/18 determined by NMR)GC-MS (METHOD 9): R_(t)=9.22 min MS: m/z=190 (fragment) (M)⁺

1H NMR (500 MHz, DMSO-d6) b 7.82-7.77 (m, 1H), 7.66-7.60 (m, 2H), 5.72(d, J=5.4 Hz, 1H), 4.59-4.55 (m, 1H), 4.39 (dd, J=9.7, 1.8 Hz, 1H), 4.23(d, J=9.7 Hz, 1H), 4.18 (dd, J=9.6, 4.1 Hz, 1H), 3.79-3.75 (d, 1H).

Step 2:

Step 2 was performed in analogy to Step 1 in the preparation ofIntermediate 38, starting from the regioisomeric mixture, to obtain[3-(2-fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester as preferred regioisomer and4-(2-Fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester (670 mg) (regioisomer ratio 80/20 determined byNMR).

GC-MS (METHOD 9): R_(t)=10.67 min

MS: m/z=265 (fragment) (M)⁺

Step 3:

Step 3 was performed in analogy to Step 3 in the preparation ofIntermediate 38, starting from the regioisomeric mixture of[3-(2-fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester and4-(2-Fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester (670 mg, 1.47 mmol), to obtain[3-(2-fluoro-4-trifluoromethyl-phenyl)-4-oxo-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester (455 mg).

GC-MS (METHOD 9): R_(t)=10.15 min

MS: m/z=249 (fragment) (M)⁺

Step 4:

Step 4 was performed in analogy to Step 4 in the preparation ofIntermediate 38, starting from[3-(2-fluoro-4-trifluoromethyl-phenyl)-4-oxo-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester (455 mg, 1.23) to obtain[3-(2-fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-4-methyl-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester as -racemic mixture (TRANS/CIS diastereoisomericratio 91/9, determined by NMR). (365 mg).

LC-MS (METHOD 10): R_(t)=3.46-3.62 min

MS (ESI pos): m/z=280 (fragment) (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b 7.64 (t, J=7.9 Hz, 1H), 7.51 (d, J=9.5 Hz,2H), 7.04 (s, 1H), 4.98 (s, 1H), 4.72-4.65 (m, 1H), 4.13 (d, J=8.4 Hz,1H), 3.94 (d, J=8.8 Hz, 1H), 3.60 (d, J=8.8 Hz, 1H), 1.31 (s, 12H).

NOE: 7.04 (NH): 3.94; 4.13; 1.31 4.98 (OH): 1.31; 4,68 1.31 (Me): 7.04;4.13; 4.98

Step 5:

Step 5 was performed in analogy to Step 5 in the preparation ofIntermediate 38, starting from[3-(2-fluoro-4-trifluoromethyl-phenyl)-4-hydroxy-4-methyl-tetrahydro-furan-3-yl]-carbamicacid tert-butyl ester (365 mg, 1.0 mmol) to obtain4-amino-4-(2-fluoro-4-trifluoromethyl-phenyl)-3-methyl-tetrahydro-furan-3-oltrifluoroacetate, as racemic mixture (TRANS/CIS diastereoisomeric ratio90/10, determined by NMR). (378 mg)

LC-MS (METHOD 1): R_(t)=2.91-3.19 min

MS (ESI pos): m/z=280 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b 8.67 (s, 3H), 7.88-7.79 (m, 2H), 7.70 (dd,J=8.5, 1.9 Hz, 1H), 5.62 (s, 1H), 4.62 (dd, J=10.0, 1.1 Hz, 1H), 4.19(dd, J=10.0, 1.5 Hz, 1H), 3.98 (d, J=9.6 Hz, 1H), 3.80 (d, J=9.6 Hz,1H), 1.48 (d, J=1.3 Hz, 3H).

Intermediate 42:

Intermediate 42 was prepared in analogy to Intermediate 35, startingfrom 4-iodobenzotrifluoride (3 g, 10.7 mmol) to obtain, afterchromatographic purification in the third step (eluent:cyclohexane/EtOAc; gradient from 0% to 100% of EtOAc), 105 mg the titlecompound, as racemic mixture (TRANS/CIS diastereoisomeric ratio 93/7,determined by NMR).

LC-MS (METHOD 5): R_(t)=0.77 min

MS (ESI pos): m/z=262 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b 7.71-7.67 (m, 2H), 7.65-7.61 (m, 2H), 4.58(d, J=5.9 Hz, 1H), 4.05 (dd, J=11.5, 1.5 Hz, 1H), 3.92-3.86 (m, 1H),3.67 (ddd, J=11.1, 4.9, 2.3 Hz, 1H), 3.57-3.52 (m, 1H), 3.48 (dq, J=5.7,1.5 Hz, 1H), 2.49-2.41 (m, 1H), 1.97 (s, 2H), 1.49-1.42 (m, 1H).

NOE 1.97 (NH2): 3.48; 4.05; 1.47 4.58 (OH): 2.46; 3.55 3.48 (CH): 1.97;3.89

Intermediate 43:

Step 1:

Step 1 was performed in analogy to Step 2 in the preparation ofIntermediate 38, starting from4-Azido-4-(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-tetrahydro-pyran-3-ol(1.7 g, 4.77 mmol) to obtain after filtration on silica, 1.3 g of thetitle compound as TRANS-racemic mixture

LC-MS (METHOD 1): R_(t)=3.93 min

MS (APCI): m/z=381 (M+H)⁺

Step 2:

To a solution of oxalylchloride (0.25 mL, 2.6 mmol) in dry DCM (20 mL)at −55° C., DMSO (0.37 mL, 0.51 mmol) was added dropwise. After 20 min,a solution of(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-3-hydroxy-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester (1.0 g, 2.37 mmol) dissolved in 3 mL of dry DCMwas added dropwise. Mixture is stirred for 1 h at −70° C., then TEA wasadded dropwise, stirred for 1.30 hrs at −40° C. and then allowed toreach room temperature. Mixture was stirred at room temperature for 48hrs. Solvent was evaporated, residue diluted with 50 ml of ethyl acetateand washed with 3×10 ml of water. Organic phase was separated, driedover sodium sulfate to obtain 0.92 g of the desired compound used in thenext step without further purification.

LC-MS (METHOD 1): R_(t)=3.79 min

MS (APCI): m/z=475 (M+H)⁺

Step 3:

Step 3 was performed in analogy to Step 4 in the preparation ofIntermediate 38, starting from4-(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-3-oxo-tetrahydro-pyran-4-yl]-carbamic acid tert-butyl ester (0.92 g, 2.41 mmol) to obtainafter chromatographic purification (eluent: cyclohexane/EtOAc; gradientfrom 100% to 40% of EtOAc), 0.190 g of CIS stereoisomer and 0.35 g ofTRANS stereoisomer, both as racemic mixture.

LC-MS (METHOD 9): R_(t)=10.49 min (CIS stereoisomer)

MS (ESI): m/z=394 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 0.88 (s, 3H) 1.22-1.38 (m, 9H) 2.26-2.42(m, 1H) 2.80 (td, J=13.54, 4.65 Hz, 1H) 3.25-3.29 (m, 1H) 3.54-3.63 (m,2H) 3.78 (br dd, J=11.37, 3.42 Hz, 1H) 5.31 (s, 1H) 6.40 (br s, 1H) 8.18(br d, J=11.61 Hz, 1H) 8.81-8.86 (m, 1H)

NOE: 6.40 (NH): 5.31 5.31 (OH): 6.40

LC-MS (METHOD 9): R_(t)=10.76 min (TRANS stereoisomer)

MS (ESI): m/z=394 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 1.03 (d, J=2.93 Hz, 3H) 1.10-1.44 (m,9H) 1.96-2.03 (m, 1H) 3.06-3.18 (m, 1H) 3.24 (d, J=11.98 Hz, 1H)3.54-3.69 (m, 1H) 3.72-3.86 (m, 2H) 4.71 (s, 1H) 7.01 (br s, 1H) 8.09(br d, J=11.74 Hz, 1H) 8.79 (s, 1H)

NOE: 7.01 (NH): 1.02; 3.62; 3.77 4.71 (OH): 3.12; 3.24

Step 4:

Step 4 was performed in analogy to Step 5 in the preparation ofIntermediate 38, starting from TRANS[(R)-4-(3-Fluoro-5-trifluoromethyl-pyridin-2-yl)-3-hydroxy-3-methyl-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester (0.350 g, 0.89 mmol) to obtain 0.25 g of desiredintermediate 42, 4-Amino-4-(3-fluoro-5-trifluoromethyl-pyridin-2-yl)-3-methyl-tetrahydro-pyran-3-ol, as trifloroacetatesalt.

LC-MS (METHOD 1): R_(t)=2.90 min

MS (APCI): m/z=295 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 1.00 (s, 3H) 1.87-1.93 (m, 1H) 3.14(ddd, J=14.49, 9.72, 4.65 Hz, 1H) 3.39-3.43 (m, 1H) 3.66-3.69 (m, 1H)3.81-3.88 (m, 1H) 4.04 (dt, J=11.86, 4.34 Hz, 1H) 5.48 (br s, 1H)8.43-8.48 (m, 1H) 8.72 (br s, 3H) 8.94 (s, 1H)

NOE: 8.72 (NH3+): 1.00; 1.90; 3.68; 3.84 1.0 (Me):8.72; 1.89; 3.66

Intermediate 44:

[4-(2-Fluoro-4-trifluoromethyl-phenyl)-3-oxo-tetrahydro-pyran-4-yl]-carbamicacid tert-butyl ester (obtained from Step 3 in the preparation ofIntermediate 38; 90 mg, 0.23 mmol) was dissolved in 1 mL of 1.4-dioxane.1.67 mL of a 1.4M solution of hydrogen chloride in 1.4-dioxane wasadded, the reaction mixture was stirred at room temperature for 1 hr andthen concentrated under vacuum. The crude obtained was stripped twicewith ethyl ether to give 70 mg of the title compound, used in the nextstep without further purification.

LC-MS (METHOD 1): R_(t)=3.70 min

MS (ESI pos): m/z=278 (M+H)⁺

EXAMPLE COMPOUNDS Example 1

To a suspension of Intermediate 5 (40.68 mg, 0.20 mmol) in 1.0 mL of dryDMF, HATU (82.47 mg, 1.3 mmol) and DIPEA (0.11 mL 0.67 mmol) were addedand the reaction mixture was stirred at room temperature. A solution ofIntermediate 13 (50 mg, 0.17 mmol) in 1.0 mL of dry DMF was added, andthe reaction mixture was stirred at room temperature overnight. Thereaction mixture was treated with basic alumina and concentrated undervacuum. The crude obtained was purified by flash chromatography (eluent:water/acetonitrile; gradient from 10% to 100% of acetonitrile) to give62 mg of the desired compound.

LC-MS (METHOD 1): R_(t)=4.78 min

MS (ESI pos): m/z=463 (M+H)⁺

The following Examples were prepared in analogy to Example 1 andpurified applying the most suitable purification technique, startingfrom the corresponding Acid and the corresponding Amine Intermediates:

MS m/z R_(t) Starting Acid Starting Amine Ex. Structure [M + H]⁺ (min)Method 2 13 2

423 4.53 METHOD 1 6 13 3

427 3.50 METHOD 10 8 14 4

403 0.90 METHOD 6 2 14 5

389 4.28 METHOD 1 5 25 6

415 4.67 METHOD 1 5 14 7

415 4.78 METHOD 1 5 15 8

431 4.98 METHOD 1 2 15 9

405 4.40 METHOD 1 5 16 10

411 0.80 METHOD 7 12 17 11

428 3.21 METHOD 10 2 17 12

388 2.86 METHOD 10 5 18 13

441 0.87 METHOD 8 5 19 14

447 3.54 METHOD 10 2 19 15

421 3.26 METHOD 10 5 20 16

415 4.70 METHDO 1 12 21 17

464 4.93 METHOD 1 8 21 18

438 3.38 METHOD 10 2 21 19

424 3.27 METHOD 10 5 21 20

450 3.59 METHOD 10 12 22 21

446 4.52 METHOD 1 8 22 22

420 3.23 METHOD 10 5 22 23

432 3.45 METHOD 10 2 22 24

406 3.12 METHDO 10 12 23 25

446 4.75 METHOD 1 8 23 26

420 3.13 METHOD 10 5 23 27

432 3.32 METHOD 10 2 23 28

406 3.01 METHOD 10 12 24 29

464 3.70 METHOD 10 5 26 30 Racemic mixture

449 3.87 METHOD 10 2 26 31 Racemic mixture

423 3.60 METHOD 10 5 27 32 Racemic mixture

[M + Na] + 453 4.78 METHOD 3 5 28 33 Racemic mixture

447 4.83 METHOD 3 5 29 34 Racemic mixture

435 4.92 METHOD 1 2 29 35 Racemic mixture

409 3.44 METHOD 10 2 30 36 Racemic mixture

391 3.36 METHOD 10 5 30 37 Racemic mixture

417 4.77 METHOD 1 5 31 38 Racemic mixture

401 4.71 METHOD 1 2 31 39 Racemic mixture

[M + Na] + 397 2.81 METHOD 4 12 32 40 Racemic mixture

450 3.63 METHOD 10 8 32 41 Racemic mixture

424 3.32 METHOD 10 12 33 42 Racemic mixture

432 3.47 METHOD 10 8 33 43 Racemic mixture

406 3.14 METHOD 10 8 34 44 Racemic mixture

406 3.25 METHOD 10 12 34 45 Racemic mixture

432 3.53 METHOD 10

Example 46

Example 46 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (34.6 mg, 0.2 mmol) and amino alcohol Intermediate 35 (65mg, 0.19 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 100% of acetonitrile),27 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=3.70 min

MS (ESI pos): m/z=439 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 2.40 (d, J=1.00 Hz, 3H) 2.59-2.73 (m,2H) 3.62-3.71 (m, 1H) 3.79-3.85 (m, 2H) 3.82 (br d, J=11.62 Hz, 2H) 4.02(br d, 1H) 4.10 (m, 1H) 5.33 (br s, 1H) 7.46-7.50 (m, 1H) 7.55 (d,J=7.99 Hz, 1H) 7.73 (t, J=8.19 Hz, 1H) 8.34 (s, 1H) 8.42 (s, 1H) 8.81(d, J=2.08 Hz, 1H) 9.19 (dd, J=1.96, 1.10 Hz, 1H)

NOE: 8.42 (NH): 4.10; 4.03; 3.65; 3.82 5.33 (OH): 4.10; 3.70

Example 47

Further elution from the column in the preparation of Example 46 gave 5mg of the title compound, as CIS— racemic mixture.

LC-MS (METHOD 1): R_(t)=3.97 min

MS (ESI pos): m/z=439 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 2.35-2.48 (m, 4H) 3.12 (br d, J=14.18Hz, 1H) 3.47 (br t, J=11.86 Hz, 1H) 3.58 (t, J=10.76 Hz, 1H) 3.72 (brdd, J=11.62, 2.57 Hz, 1H) 3.74-3.84 (m, 1H) 4.10 (dd, J=10.03, 5.14 Hz,1H) 5.66 (s, 1H) 7.47-7.59 (m, 2H) 7.61-7.67 (m, 1H) 8.49 (s, 1H) 8.62(s, 1H) 8.76 (d, J=1.96 Hz, 1H) 9.21 (dd, J=2.08, 1.10 Hz, 1H)

NOE: 8.62 (NH): 4.10; 5.66; 2.49; 3.78; 3.12 5.66(OH): 8.62; 4.10; 3.78;3.12

The following Examples were prepared in analogy to Example 46 andExample 47 starting from the corresponding acid and amino alcoholIntermediates:

Starting Amino- MS m/z R_(t) Starting Acid alcohol Ex. Structure [M +H]⁺ (min) Method 5 35 48 TRANS- Racemic mixture

465 4.12 METHOD 1 5 35 49 CIS- Racemic mixture

465 4.33 METHOD 1

Relative stereochemistry assigned by NMR:

Relative Example stereochemistry 1H-NMR NOE 48

TRANS Racemic mixture 1H NMR (500 MHz, DMSO-d6) δ ppm 0.94-1.11 (m, 4 H)2.11 (tt, J = 8.47, 5.10 Hz, 1 H) 2.56-2.76 (m, 2 H) 3.66 (td, J =11.68, 2.08 Hz, 1 H) 3.81 (br d, J = 11.25 Hz, 2 H) 4.03 (d, J = 11.74Hz, 1 H) 4.09 (m, 1 H) 5.33 (br s, 1 H) 7.46-7.50 (m, 1 H) 7.54 (d, J =8.03 Hz, 1 H) 7.73 (t, J = 7.98 Hz, 1 H) 8.33 (s, 1 H) 8.40 (s, 1 H)8.77 (d, 8.40 (NH): 4.09 5.33 (OH): 4.09 J = 2.20 Hz, 1 H) 9.13 (d, J =2.20 Hz, 1 H) 49

CIS-Racemic mixture 1H NMR (500 MHz, DMSO-d6) δ ppm 0.86-1.07 (m, 4 H)2.06- 2.16 (m, 1 H) 2.42- 2.48 (m, 1 H) 3.12 (br d, J = 14.18 Hz, 1 H)3.26- 3.29 (m, 1 H) 3.47 (br t, J = 11.98 Hz, 1 H) 3.58 (t, J = 10.76Hz, 1 H) 3.65-3.75 (m, 1 H) 3.78 (dd, J = 11.49, 5.14 Hz, 1 H) 4.09 (dd,J = 10.27, 5.14 Hz, 1 H) 5.66 (br s, 1 H) 7.48-7.60 (m, 8.58 (NH): 5.66;3.58; 3.47; 3.12 5.66 (OH): 8.58; 3.77 4.09 (CH): 2.46 2 H) 7.60-7.65(m, 1 H) 8.48 (s, 1 H) 8.58 (s, 1 H) 8.71 (d, J = 2.20 Hz, 1 H) 9.14 (d,J = 1.71 Hz, 1 H)

Example 50

Example 50 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (100 mg, 0.56 mmol) and amino alcohol Intermediate 36(171.4 mg, 0.62 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 0% to 60% of acetonitrile),209 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=3.04 min

MS (ESI pos): m/z=422 (M+H)⁺

1H NMR (400 MHz, DMSO-d6) b ppm 2.41 (s, 3H) 2.45-2.48 (m, 1H) 2.64-2.74(m, 1H) 3.63-3.75 (m, 1H) 3.76-3.87 (m, 2H) 3.94 (br d, J=4.89 Hz, 1H)4.08 (d, J=11.74 Hz, 1H) 5.30 (d, J=5.67 Hz, 1H) 7.83 (d, J=8.22 Hz, 1H)8.10 (dd, J=8.31, 1.66 Hz, 1H) 8.38 (s, 1H) 8.44 (s, 1H) 8.82 (d, J=1.96Hz, 1H) 8.85 (d, J=1.76 Hz, 1H) 9.20 (dd, J=1.96, 0.98 Hz, 1H)

NOE: 8.44 (NH): 4.08; 3,94; 3.68; 2.48 5.32(OH): 8.58; 3,78, 2.68 3.94(CH): 8.44; 2.48

The following Examples were prepared in analogy to Example 50 startingfrom the corresponding acid and amino alcohol Intermediates:

Starting Starting Amino- MS m/z R_(t) Acid alcohol Ex. Structure [M +H]⁺ (min) Method 12 36 51 TRANS- Racemic mixture

462 3.87 METHOD 1  5 36 52 TRANS- Racemic mixture

448 3.58 METHOD 1

Relative stereochemistry assigned by NMR:

Relative Example stereochemistry 1H-NMR 51

TRANS Racemic mixture 1H NMR (400 MHz, DMSO-d6) δ ppm 0.86-1.07 (m, 4 H)2.03- 2.17 (m, 1 H) 2.47 (s, 3 H) 2.49- 2.5 (m, 1H) 2.68 (td, J = 12.57,4.40 Hz, 1 H) 3.64-3.72 (m, 1 H) 3.77-3.88 (m, 3 H) 4.10 (d, J = 12.13Hz, 1 H) 5.28 (d, J = 5.67 Hz, 1 H) 7.83 (d, J = 8.22 Hz, 1 H) 8.08 (d,J = 7.73 Hz, 1 H) 8.65 (s, 1 H) 8.71 (d, J = 1.96 Hz, 1 H) 8.83 (s, 1 H)9.03 (d, J = 1.96 Hz, 1 H) 52

TRANS Racemic mixture 1H NMR (400 MHz, DMSO-d6) δ ppm 0.91-1.09 (m, 4 H)2.08- 2.15 (m, 1 H) 2.46 (s, 1 H) 2.59- 2.78 (m, 1 H) 3.64-3.75 (m, 1 H)3.76-3.86 (m, 2 H) 3.92 (br d, J = 4.21 Hz, 1 H) 4.08 (d, J = 11.93 Hz,1 H) 5.29 (d, J = 5.77 Hz, 1 H) 7.83 (d, J = 8.31 Hz, 1 H) 8.10 (dd, J =8.12, 2.15 Hz, 1 H) 8.37 (s, 1 H) 8.43 (s, 1 H) 8.77 (d, J = 2.15 Hz, 1H) 8.84 (d, J = 1.96 Hz, 1 H) 9.14 (d, J = 2.15 Hz, 1 H)

Example 53

Example 53 was synthesized in analogy to Example 1, starting from acidIntermediate 5 (98 mg, 0.48 mmol) and amino alcohol Intermediate 38 (198mg, 0.48 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 80% of acetonitrile),7 mg of the title compound, as CIS-racemic mixture.

LC-MS (METHOD 1): R_(t)=4.45 min

MS (ESI pos): m/z=479 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 0.94-1.07 (m, 4H) 1.12 (s, 3H) 2.12 (tt,J=8.38, 5.20 Hz, 1H) 2.72 (br t, J=11.98 Hz, 1H) 2.98 (br d, J=13.69 Hz,1H) 3.47 (d, J=11.25 Hz, 1H) 3.56 (br t, J=11.74 Hz, 1H) 3.72-3.76 (m,1H) 3.76-3.87 (m, 1H) 5.38 (s, 1H) 7.46-7.56 (m, 2H) 7.70 (t, J=8.07 Hz,1H) 8.35 (s, 1H) 8.72 (d, J=2.20 Hz, 1H) 8.88-8.95 (m, 1H) 9.12 (d,J=2.20 Hz, 1H)

NOE: 8.92 (NH): 5.38; 3,80 5.38(OH): 8.92; 3,80

Example 54

Further elution from the column in the preparation of Example 53 gave 29mg of the title compound, as racemic mixture (TRANS/CISdiastereoisomeric ratio 92/8 determined by NMR).

LC-MS (METHOD 1): R_(t)=4.45 min

MS (ESI pos): m/z=479 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 0.95-1.07 (m, 4H) 1.10 (d, 3H) 2.12 (tt,J=8.44, 5.14 Hz, 1H) 2.51-2.56 (m, 1H) 3.03 (td, J=12.65, 4.28 Hz, 1H)3.53-3.65 (m, 2H) 3.74-3.76 (m, 1H) 3.88 (d, J=12.23 Hz, 1H) 5.10 (s,1H) 7.44-7.48 (dd, 1H) 7.53 (d, J=8.27 Hz, 1H) 7.66 (t, J=8.05 Hz, 1H)8.38 (s, 1H) 8.48 (s, 1H) 8.79 (d, J=2.20 Hz, 1H) 9.16 (d, J=2.20 Hz,1H)

NOE: 8.48 (NH): 1.09; 3,88; 2.53 5.10(OH): 3,04, 3.57 1.09 (Me): 8.48;3,57; 2.53; 3.88

Example 55

Example 55 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (304 mg, 1.68 mmol) and amino alcohol Intermediate 38(700 mg, 1.68 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 100% of acetonitrile),300 mg of the title compound, as racemic mixture (TRANS/CISdiastereoisomeric ratio 96/4 determined by NMR).

LC-MS (METHOD 1): R_(t)=4.02 min

MS (ESI pos): m/z=453 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 1.10 (d, J=1.00 Hz, 3H) 2.41 (d, J=0.86Hz, 3H) 2.52-2.58 (m, 1H) 3.03 (br d, J=4.16 Hz, 1H) 3.46-3.66 (m, 2H)3.72-3.82 (m, 1H) 3.88 (d, J=12.23 Hz, 1H) 5.10 (s, 1H) 7.46 (br d,J=12.72 Hz, 1H) 7.50-7.57 (m, 1H) 7.67 (t, J=8.23 Hz, 1H) 8.35-8.39 (m,1H) 8.50 (s, 1H) 8.83 (d, J=1.96 Hz, 1H) 9.18-9.24 (m, 1H)

NOE: 8.50 (NH): 1.10; 3,89; 3.64 5.10(OH): 3,03, 3.58 1.10 (Me): 8.50;3,56; 3.89

Example 56

Example 56 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (36 mg, 0.21 mmol) and amino alcohol Intermediate 39 (55mg, 0.2 mmol) to give, after flash chromatographic purification (eluent:water/acetonitrile; gradient from 10% to 65% of acetonitrile), 52 mg ofthe title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=3.57 min

MS (ESI pos): m/z=440 (M+H)⁺

1H NMR (400 MHz, DMSO-d6) b ppm 2.40 (s, 3H) 2.42-2.48 (m, 1H) 2.89 (td,J=13.35, 4.40 Hz, 1H) 3.65 (br t, J=11.30 Hz, 1H) 3.79-3.89 (m, 2H) 4.01(d, J=12.42 Hz, 1H) 4.09 (s, 1H) 5.35 (br s, 1H) 8.11 (d, J=11.84 Hz,1H) 8.35 (s, 1H) 8.46 (s, 1H) 8.81 (d, J=1.76 Hz, 2H) 9.20 (s, 1H)

Example 57

Example 57 was synthesized in analogy to Example 1, starting from acidIntermediate 5 (42 mg, 0.21 mmol) and amino alcohol Intermediate 39 (55mg, 0.2 mmol) to give, after flash chromatographic purification (eluent:water/acetonitrile; gradient from 10% to 70% of acetonitrile), 48 mg ofthe title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=4.00 min

MS (ESI pos): m/z=466 (M+H)⁺

1H NMR (400 MHz, DMSO-d6) b ppm 0.87-1.11 (m, 4H) 2.07-2.15 (m, 1H) 2.47(m, 1H) 2.89 (td, J=13.11, 4.11 Hz, 1H) 3.64 (br t, J=11.54 Hz, 1H)3.78-3.89 (m, 2H) 4.01 (d, J=12.52 Hz, 1H) 4.09 (br s, 1H) 5.35 (br s,1H) 8.11 (d, J=11.93 Hz, 1H) 8.34 (s, 1H) 8.44 (s, 1H) 8.77 (s, 1H) 8.81(s, 1H) 9.14 (d, J=1.96 Hz, 1H)

Example 58

Example 58 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (187 mg, 1.06 mmol) and amino alcohol Intermediate 41(378 mg, 0.96 mmol) to give, after flash chromatographic purification(eluent: dichloromethane/MeOH; gradient from 0% to 60% of MeCOH), 22 mgof the title compound, as racemic mixture (TRANS/CIS diastereoisomericratio 91/9)

LC-MS (METHOD 10): R_(t)=3.16 min

MS (ESI pos): m/z=439 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 1.56 (s, 3H) 2.40 (d, J=0.73 Hz, 4H)3.78 (d, J=9.05 Hz, 1H) 3.90 (d, J=9.05 Hz, 1H) 4.39 (d, J=8.80 Hz, 1H)4.90 (d, J=8.80 Hz, 1H) 5.31 (s, 1H) 7.48-7.56 (m, 2H) 7.71 (t, J=8.07Hz, 1H) 8.37 (s, 1H) 8.67 (s, 1H) 8.82 (d, J=1.96 Hz, 1H) 9.09-9.26 (m,1H) 9.21 (dd, J=2.08, 1.10 Hz, 1H)

NOE: 8.67 (NH): 1.56; 3,90; 4.38 5.31 (OH): 3,78, 4.90 1.56 (Me): 8.67

Example 59

Example 59 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (35.6 mg, 0.20 mmol) and amino alcohol Intermediate 42(50 mg, 0.19 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 65% of acetonitrile),42 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=3.77 min

MS (ESI pos): m/z=421 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 2.40 (d, J=0.98 Hz, 3H) 2.51-2.54 (m,1H) 2.62-2.70 (m, 1H) 3.67 (td, J=11.68, 1.83 Hz, 1H) 3.75-3.82 (m, 2H)3.82-3.89 (m, 1H) 4.06 (d, J=11.49 Hz, 1H) 5.14 (br d, J=4.16 Hz, 1H)7.62-7.70 (m, 4H) 8.35 (s, 1H) 8.37 (s, 1H) 8.80 (d, J=1.96 Hz, 1H) 9.19(dd, J=2.08, 1.10 Hz, 1H)

Example 60

Example 60 was synthesized in analogy to Example 1, starting from acidIntermediate 5 (40.8 mg, 0.2 mmol) and amino alcohol Intermediate 42 (50mg, 0.19 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 70% of acetonitrile),39 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=4.07 min

MS (ESI pos): m/z=424 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) δ ppm 0.86-1.06 (m, 4H) 2.07-2.14 (m, 1H)2.51-2.53 (m, 1H) 2.62-2.70 (m, 1H) 3.66 (td, J=11.74, 1.71 Hz, 1H)3.75-3.87 (m, 3H) 4.07 (d, J=11.25 Hz, 1H) 5.14 (d, J=5.62 Hz, 1H)7.62-7.70 (m, 4H) 8.35 (s, 1H) 8.36 (s, 1H) 8.76 (d, J=2.20 Hz, 1H) 9.13(d, J=2.20 Hz, 1H)

Example 61

Example 61 was synthesized in analogy to Example 1, starting from acidIntermediate 12 (82 mg, 0.29 mmol) and amino alcohol Intermediate 39(66.7 mg, 0.31 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 70% of acetonitrile),110 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=4.22 min

MS (ESI pos): m/z=480 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 0.90-1.06 (m, 4H) 2.09 (tt, J=8.44, 5.14Hz, 1H) 2.45 (s, 3H) 2.47 (m, 1H) 2.88 (td, J=13.27, 4.28 Hz, 1H)3.59-3.67 (m, 1H) 3.77-3.88 (m, 2H) 4.01-4.05 (m, 1H) 4.08 (br d, J=5.14Hz, 1H) 5.32 (d, J=5.62 Hz, 1H) 8.11 (dd, J=11.86, 1.59 Hz, 1H) 8.69 (s,1H) 8.71 (d, J=2.36 Hz, 1H) 8.81 (s, 1H) 9.03 (d, J=2.20 Hz, 1H)

NOE: 8.69 (NH): 4.08; 2.47 3.64 5.32(OH): 3,82, 2.88 4.08 (CH): 8.69;2.47

Example 62

Example 62 was synthesized in analogy to Example 1, starting from acidIntermediate 12 (103 mg, 0.6 mmol) and amino alcohol Intermediate 43(250 mg, 0.6 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 70% of acetonitrile),115 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=4.40 min

MS (ESI pos): m/z=494 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 0.92-1.06 (m, 4H) 1.21 (d, J=3.18 Hz,3H) 2.10 (tt, J=8.47, 5.23 Hz, 1H) 2.30-2.36 (m, 1H) 2.47-2.49 (m, 3H)3.22-3.29 (m, 1H) 3.49-3.54 (m, 1H) 3.62 (br t, J=10.88 Hz, 1H)3.76-3.88 (m, 2H) 4.96 (s, 1H) 8.08 (dd, J=1 1.98, 1.47 Hz, 1H) 8.72 (d,J=1.96 Hz, 1H) 8.78 (s, 1H) 8.84 (s, 1H) 9.05 (d, J=2.20 Hz, 1H)

NOE: 8.78 (NH): 1.21; 2.34; 3.62 4.96(OH): 3.51, 3.35 1.21 (Me): 8.78;2.34

Example 63

Example 63 was synthesized in analogy to Example 1, starting from acidIntermediate 5 (45 mg, 0.22 mmol) and amino alcohol Intermediate 40 (55mg, 0.21 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 70% of acetonitrile),55 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=3.82 min

MS (ESI pos): m/z=448 (M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 0.91-1.08 (m, 4H) 2.06-2.17 (m, 1H)2.41-2.48 (m, 1H) 2.76-2.84 (m, 1H) 3.65-3.76 (m, 2H) 3.92 (dt, J=11.23,3.58 Hz, 1H) 3.95-4.00 (m, 1H) 4.04 (br s, 1H) 5.24 (d, J=4.03 Hz, 1H)7.75 (d, J=8.44 Hz, 1H) 8.11 (dd, J=8.50, 2.02 Hz, 1H) 8.33 (s, 1H) 8.43(s, 1H) 8.76 (d, J=2.08 Hz, 1H) 8.88 (dd, J=1.53, 0.79 Hz, 1H) 9.13 (d,J=2.20 Hz, 1H)

NOE 8.43 (NH): 4.04; 2.44; 3.98 5.24(OH): 3.66, 2.80 4.04 (CH): 8.43;2.44

Example 64

Example 64 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (39 mg, 0.22 mmol) and amino alcohol Intermediate 40 (55mg, 0.21 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 65% of acetonitrile),31 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 10): R_(t)=2.66 min

MS (ESI pos): m/z=422(M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 2.40 (s, 3H) 2.42 (br s, 1H) 2.76-2.85(m, 1H) 3.64-3.77 (m, 2H) 3.90-3.95 (m, 1H) 3.97 (br d, J=12.23 Hz, 1H)4.05 (br s, 1H) 5.24 (d, J=5.56 Hz, 1H) 7.75 (d, J=8.44 Hz, 1H) 8.11(dd, J=8.47, 2.11 Hz, 1H) 8.33 (s, 1H) 8.45 (s, 1H) 8.80 (d, J=1.90 Hz,1H) 8.88 (s, 1H) 9.19 (s, 1H)

NOE: 8.45 (NH): 4.05; 2.44; 3.98; 3.68 5.24(OH): 3.72; 3.97; 2.80 4.05(CH): 8.45; 2.44

Example 65

Example 65 was synthesized in analogy to Example 1, starting from acidIntermediate 12 (78 mg, 0.36 mmol) and amino alcohol Intermediate 40 (90mg, 0.34 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 10% to 70% of acetonitrile),110 mg of the title compound, as TRANS-racemic mixture.

LC-MS (METHOD 1): R_(t)=4.07 min

MS (ESI pos): m/z=462(M+H)⁺

1H NMR (500 MHz, DMSO-d6) b ppm 0.90-1.07 (m, 4H) 2.09 (tt, J=8.59, 4.98Hz, 1H) 2.42-2.47 (m, 4H) 2.79 (ddd, J=13.66, 11.34, 4.34 Hz, 1H)3.64-3.76 (m, 2H) 3.91 (dt, J=11.28, 3.59 Hz, 1H) 3.99 (d, J=12.35 Hz,1H) 3.98-3.99 (m, 1H) 4.02 (br s, 1H) 5.21 (d, J=4.52 Hz, 1H) 7.73 (d,J=8.44 Hz, 1H) 8.12 (dd, J=8.50, 2.14 Hz, 1H) 8.69 (d, J=10.11 Hz, 2H)8.88 (dd, J=1.47, 0.73 Hz, 1H) 9.02 (d, J=1.83 Hz, 1H)

Example 66

Example 66 was synthesized in analogy to Example 1, starting from acidIntermediate 5 (16.8 mg, 0.08 mmol) and amino alcohol Intermediate 44(20 mg, 0.08 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 0% to 100% of acetonitrile),11 mg of the title compound.

LC-MS (METHOD 1): R_(t)=4.78 min

MS (ESI pos): m/z=463 (M+H)⁺

Example 67

Example 67 was synthesized in analogy to Example 1, starting from acidIntermediate 2 (19.7 mg, 0.11 mmol) and amino alcohol Intermediate 44(35 mg, 0.11 mmol) to give, after flash chromatographic purification(eluent: water/acetonitrile; gradient from 0% to 100% of acetonitrile),24 mg of the title compound.

LC-MS (METHOD 1): R_(t)=4.40 min

MS (ESI pos): m/z=437 (M+H)⁺

Examples 68, 69, 70, 71

Examples 68, 69, 70 and 71 were synthesized in analogy to Example 1,starting from acid Intermediate 5 (150 mg, 0.74 mmol) and amino alcoholIntermediate 37 (230 mg, 0.71 mmol) to give, after flash chromatographicpurification (eluent: water/acetonitrile; gradient from 0% to 80% ofacetonitrile), 143 mg of mixture of the title compounds, which wereobtained as single stereoisomers by chiral HPLC separation.

MS R_(t) (min) R_(t) (min) m/z [LC-MS [Chiral HPLC Ex. # Structure [M +H]⁺ Method] Method] 68 CIS single stereoisomer a

451 4.28 METHOD 1  9.66 [C3] 69 CIS single stereoisomer b

451 4.28 METHOD 1 11.30 [C3] 70 TRANS single stereoisomer a

451 4.21 METHOD 1  16.32, [C3] 71 TRANS single stereoisomer b

451 4.23 METHOD 1 22.71 [C3]

Relative stereochemistry assigned by NMR:

Relative Example stereochemistry 1H-NMR NOE 68

CIS single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 0.94-1.06 2.14(m, 1 H) 3.58 (dd, J = 9.29, 5.14 Hz, 1 H) 4.02 (dd, J = 9.29, 5.87 Hz,1 H) 4.14 (d, J = 9.78 Hz, 1 H) 4.40 (br s, 1 H) 4.80 (d, J = 10.03 Hz,1 H) 6.34 (br s, 1 H) 7.54-7.63 (m, 2 H) 7.63-7.68 (m, 1 H) 8.39 (s, 1H) 8.72 (d, J = 2.20 Hz, 1 H) 9.09-9.13 (m, 2 H) 9.11 (NH): 6.34; 4.14;3.58 6.34(OH): 9.11, 4.14; 3.58 4.40 (CH): 4.80 69

CIS single stereoisomer b 1H NMR (500 MHz, DMSO-d6) δ ppm 0.94-1.06 (m,4 H) 2.07- 2.14 (m, 1 H) 3.58 (dd, J = 9.29, 5.14 Hz, 1 H) 4.02 (dd, J =9.29, 5.87 Hz, 1 H) 4.14 (d, J = 9.78 Hz, 1 H) 4.40 (br s, 1 H) 4.80 (d,J = 10.03 Hz, 1 H) 6.34 (br s, 1 H) 7.54-7.63 (m, 2 H) 7.63-7.68 (m, 1H) 8.39 (s, 1 H) 8.72 (d, J = 2.20 Hz, 1 H) 9.09-9.13 (m, 2 H) 9.11(NH): 6.34; 4.14; 3.58 6.34(OH): 9.11, 4.14; 3.58 4.40 (CH): 4.80 70

TRANS single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 0.92-1.07(m, 4 H) 3.71- 3.80 (m, 1 H) 4.25 (dd, J = 9.78, 4.40 Hz, 1 H) 4.35 (d,J = 8.80 Hz, 1 H) 4.69 (d, J = 8.80 Hz, 1 H) 4.76 (br s, 1 H) 5.64 (brs, 1 H) 7.51-7.56 (m, 2 H) 7.73 (t, J = 7.82 Hz, 1 H) 8.28 (s, 1 H) 8.36(s, 1 H) 8.74 (d, J = 1.96 Hz, 1 H) 9.09-9.11 (m, 1 H) 8.28 (NH): 4.69;4.76; 4.25 5.64(OH): 4.35; 3.76 4.76 (CH): 8.28 71

TRANS single stereoisomer b 1H NMR (500 MHz, DMSO-d6) δ ppm 0.92-1.07(m, 4 H) 3.71- 3.80 (m, 1 H) 4.25 (dd, J = 9.78, 4.40 Hz, 1 H) 4.35 (d,J = 8.80 Hz, 1 H) 4.69 (d, J = 8.80 Hz, 1 H) 4.76 (br s, 1 H) 5.64 (brs, 1 H) 7.51-7.56 (m, 2 H) 7.73 (t, J = 7.82 Hz, 1 H) 8.28 (s, 1 H) 8.36(s, 1 H) 8.74 (d, J = 1.96 Hz, 1 H) 9.09-9.11 (m, 1 H) 8.28 (NH): 4.69;4.76; 4.25 5.64(OH): 4.35; 3.76 4.76 (CH): 8.28

Examples 72, 73, 74, 75

Examples 72, 73, 74 and 75 were synthesized in analogy to Example 1,starting from acid Intermediate 2 (92 mg, 0.52 mmol) and amino alcoholIntermediate 37 (160 mg, 43% content, 0.26 mmol) to give, after twosubsequent flash chromatographic purifications (eluent:water/acetonitrile; gradient from 0% to 80% of acetonitrile; eluent:DCM/isopropyl alcohol; gradient from 0% to 30% of isopropyl alcohol),110 mg of mixture of the title compounds, which were obtained as singlestereoisomers by chiral HPLC separation.

MS R_(t) (min) R_(t) (min) m/z [LC-MS [Chiral HPLC Ex. # Structure [M +H]⁺ Method] Method] 72 CIS single stereoisomer a

425 3.84 METHOD 1 12.08 [C3] 73 CIS single stereoisomer b

425 3.85 METHOD 1 13.41 [C3] 74 TRANS single stereoisomer a

425 3.75 METHOD 1 22.54 [C3] 75 TRANS single stereoisomer b

425 3.78 METHOD 1 26.59 [C3]

The following examples were obtained as single stereoisomers by chiralHPLC separation of the corresponding racemic mixture:

Starting Racemic MS R_(t) (min) R_(t) (min) Mixture m/z [LC-MS [ChiralHPLC Ex. # Ex. # Structure [M + H]⁺ Method] Method] 40 80 a Singlestereoisomer a

450 3.89 METHOD 10  5.98 [C1] 40 80b single stereoisomer b

450 3.89 METHOD 10  6.76 [C1] 46 TRANS- Racemic mixture 81a TRANS-single stereoisomer a

439 3.67 METHOD 1 11.19 [C4] 46 TRANS- Racemic mixture 81b TRANS- singlestereoisomer b

439 3.67 METHOD 1 13.99 [C4] 48 TRANS- Racemic mixture 82a TRANS- singlestereoisomer a

465 3.41 METHOD 10 11.78 [C4] 48 TRANS- Racemic mixture 82b TRANS-single stereoisomer b

465 3.41 METHOD 10 14.22 [C4] 55 TRANS/ CIS 96/4 Racemic mixture 83aTRANS- single stereoisomer a

453 METHOD 1 11.71 [C3] 55 TRANS/ CIS 96/4 Racemic mixture 83b TRANS-single stereoisomer b

453 4.07 METHOD 1 26.90 [C3] 55 TRANS/ CIS 96/4 Racemic mixture 84a CIS-single stereoisomer a

453 4.07 METHOD 1 13.67 [C3] 55 TRANS/ CIS 96/4 Racemic mixture 84b CIS-single stereoisomer b

453 4.07 METHOD 1 18.13 [C3] 54 TRANS/ CIS 92/8 Racemic mixture 85a CIS-single stereoisomer a

479 4.48 METHOD 1  5.46 [C1] 54 TRANS/ CIS 92/8 Racemic mixture 85b CIS-single stereoisomer b

479 4.48 METHOD 1  6.73 [C1] 54 TRANS/ CIS 92/8 Racemic mixture 86aTRANS- single stereoisomer a

479 4.48 METHOD 1  6.08 [C1] 54 TRANS/ CIS 92/8 Racemic mixture 86bTRANS- single stereoisomer b

479 4.48 METHOD 1 10.75 [C1] 50 TRANS- Racemic mixture 87a TRANS- singlestereoisomer a

422 3.08 METHOD 1  9.97 [C6] 50 TRANS- Racemic mixture 87b TRANS- singlestereoisomer b

422 3.08 METHOD 1 12.98 [C6] 51 TRANS- Racemic mixture 88a TRANS- singlestereoisomer a

462 3.87 METHOD 1  7.71 [C1] 51 TRANS- Racemic mixture 88b TRANS- singlestereoisomer b

462 3.87 METHOD 1  9.49 [C1] 52 TRANS- Racemic mixture 89a TRANS- singlestereoisomer a

448 3.60 METHOD 1 11.82 [C6] 52 TRANS- Racemic mixture 89b TRANS- singlestereoisomer b

448 3.60 METHOD 1 16   [C6] 61 TRANS- Racemic mixture 90a TRANS singlestereoisomer a

480 4.22 METHOD 1  4.9  [C7] 61 TRANS- Racemic mixture 90b TRANS singlestereoisomer b

480 4.22 METHOD 1  5.54 [C7] 62 TRANS- Racemic mixture 91a TRANS singlestereoisomer a

494 4.4  METHOD 1  6.68 [C1] 62 TRANS- Racemic mixture 91b TRANS singlestereoisomer b

494 4.4  METHOD 1 12.94 [C1] 58 TRANS/ CIS 91/9 Racemic mixture 92a CISSingle stereoisomer a

439 3.16 METHOD 10  4.76 [C1] 58 TRANS/ CIS 91/9 Racemic mixture 92b CISSingle stereoisomer b

439 3.16 METHOD 10  8.24 [C1] 58 TRANS/ CIS 91/9 Racemic mixture 93aTRANS Single stereoisomer a

439 3.16 METHOD 10  7.20 [C1] 58 TRANS/ CIS 91/9 Racemic mixture 93bTRANS Single stereoisomer b

439 3.16 METHOD 10 16.60 [C1]

Relative stereochemistry assigned by NMR:

Relative Example stereochemistry 1H-NMR NOE 81a

TRANS-single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 2.40 (s, 3H) 2.60-2.71 (m, 2 H) 3.63-3.69 (m, 1 H) 3.82 (br d, J = 11.98 Hz, 2 H)4.03 (d, J = 11.74 Hz, 1 H) 4.08- 4.11 (m, 1 H) 5.32 (d, J = 5.38 Hz, 1H) 7.46-7.56 (m, 2 H) 7.73 (t, J = 7.51 Hz, 1 H) 8.34 (s, 1 H) 8.42 (s,1 H) 8.81 (d, J = 2.20 Hz, 1 H) 9.17-9.19 (m, 1 8.42 (NH): 4.08; 4.03;3.65; 3.82 5.32 (OH): 4.10; 3.70. 82a

TRANS-single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 0.93-1.06(m, 4 H) 2.10 (tt, J = 8.38, 5.20 Hz, 1 H) 2.59-2.71 (m, 2 H) 3.66 (td,J = 11.68, 2.08 Hz, 1 H) 3.81 (br d, J = 11.25 Hz, 2 H) 4.03 (d, J =11.86 Hz, 1 H) 4.09 (br d, J = 4.65 Hz, 1 H) 5.32 (d, J = 5.38 Hz, 1 H)7.48 (d, J = 12.47 Hz, 1 H) 7.54 (dd, J = 8.31, 1.22 Hz, 1 H) 7.73 (t, J= 8.19 Hz, 1 H) 8.40 (NH): 4.09 5.32(OH): 4.09 8.33 (s, 1 H) 8.40 (s, 1H) 8.77 (d, J = 2.20 Hz, 1 H) 9.13 (s, 1 H) 83a

TRANS-single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 1.10 (d, J =2.57 Hz, 3 H) 2.41 (d, J = 0.73 Hz, 3 H) 2.52-2.57 (m, 1 H) 3.03 (br d,J = 4.40 Hz, 1 H) 3.56-3.58 (m, 1 H) 3.60-3.65 (m, 1 H) 3.75 (br dd, J =11.49, 2.57 Hz, 1 H) 3.88 (d, J = 12.23 Hz, 1 H) 5.10 (s, 1 H) 7.46 (dd,J = 12.78, 1.41 Hz, 1 H) 7.53 (dd, 8.50 (NH): 1.10; 3.88; 2.53 5.10(OH):3.57, 3.03 1.10 (Me): 8.50; 3.88; 2.53 J = 8.31, 1.34 Hz, 1 H) 7.67 (t,J = 8.25 Hz, 1 H) 8.39 (s, 1 H) 8.50 (s, 1 H) 8.83 (d, J = 2.08 Hz, 1 H)9.23 (dd, J = 1.96, 1.10 Hz, 1 H) 84A

CIS-single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 1.12 (s, 2 H)2.36-2.47 (m, 3 H) 2.67-2.84 (m, 1 H) 2.98 (br d, J = 14.18 Hz, 1 H)3.47 (d, J = 11.00 Hz, 1 H) 3.56 (br t, J = 11.98 Hz, 1 H) 3.75 (br dd,J = 11.37, 3.55 Hz, 1 H) 3.81 (d, J = 11.25 Hz, 1 H) 5.39 (s, 1 H) 7.46-7.55 (m, 2 H) 7.70 (t, J = 8.07 Hz, 1 H) 8.96 (NH): 5.39; 2.73 5.39(OH):8.96; 2.73 8.35 (s, 1 H) 8.77 (d, J = 1.96 Hz, 1 H) 8.96 (s, 1 H) 9.18(dd, J = 2.08, 1.10 Hz, 1 H) 84b

CIS-single stereoisomer b 1H NMR (500 MHz, DMSO-d6) δ ppm 1.12 (s, 2 H)2.36-2.47 (m, 3 H) 2.67-2.84 (m, 1 H) 2.98 (br d, J = 14.18 Hz, 1 H)3.47 (d, J = 11.00 Hz, 1 H) 3.56 (br t, J = 11.98 Hz, 1 H) 3.75 (br dd,J = 11.37, 3.55 Hz, 1 H) 3.81 (d, J = 11.25 Hz, 1 H) 5.39 (s, 1 H) 7.46-7.55 (m, 2 H) 7.70 (t, J = 8.07 Hz, 1 H) 8.96 (NH): 5.39; 2.73 5.39(OH):8.96; 2.73 8.35 (s, 1 H) 8.77 (d, J = 1.96 Hz, 1 H) 8.96 (s, 1 H) 9.18(dd, J = 2.08, 1.10 Hz, 1 H) 83b

TRANS-single stereoisomer b 1H NMR (500 MHz, DMSO-d6) δ ppm 1.10 (d, J =2.57 Hz, 3 H) 2.41 (d, J = 0.73 Hz, 3 H) 2.52-2.57 (m, 1 H) 3.03 (br d,J = 4.40 Hz, 1 H) 3.56-3.58 (m, 1 H) 3.60-3.65 (m, 1 H) 3.75 (br dd, J =11.49, 2.57 Hz, 1 H) 3.88 (d, J = 12.23 Hz, 1 H) 5.10 (s, 1 H) 7.46 (dd,J = 12.78, 1.41 Hz, 1 H) 7.53 (dd, 8.50 (NH): 1.10; 3.88; 2.53 5.10(OH):3.57, 3.03 1.10 (Me): 8.50; 3.88; 2.53 J = 8.31, 1.34 Hz, 1 H) 7.67 (t,J = 8.25 Hz, 1 H) 8.39 (s, 1 H) 8.50 (s, 1 H) 8.83 (d, J = 2.08 Hz, 1 H)9.23 (dd, J = 1.96, 1.10 Hz, 1 H) 85a

CIS-single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 0.94-1.07 (m,4 H) 1.12 (s, 3 H) 2.12 (tt, J = 8.38, 5.20 Hz, 1 H) 2.72 (br t, J =11.98 Hz, 1 H) 2.98 (br d, J = 13.69 Hz, 1 H) 3.47 (d, J = 11.25 Hz, 1H) 3.56 (br t, J = 11.74 Hz, 1 H) 3.75 (br dd, J = 11.37, 3.30 Hz, 1 H)3.81 (d, J = 11.25 Hz, 1 H) 5.38 (s, 1 H) 7.46- 7.56 (m, 2 H) 7.70 (t, J= 8.07 Hz, 1 H) 8.92 (NH): 5.38; 3.81; 3.56 5.38(OH): 8.92; 1.12 (Me):3.47; 2.72 8.35 (s, 1 H) 8.72 (d, J = 2.20 Hz, 1 H) 8.92 (s, 1 H) 9.12(d, J = 2.20 Hz, 1 H) 86a

TRANS-single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 0.95-1.07(m, 5 H) 1.10 (d, J = 2.57 Hz, 2 H) 2.12 (tt, J = 8.44, 5.14 Hz, 1 H)2.51- 2.56 (m, 1 H) 3.03 (td, J = 12.65, 4.28 Hz, 1 H) 3.54- 3.59 (m, 1H) 3.62 (t, J = 11.25 Hz, 1 H) 3.75 (br dd, J = 11.25, 2.45 Hz, 1 H)3.88 (d, J = 12.23 Hz, 1 H) 5.10 (s, 1 H) 7.46 (br d, J = 12.96 Hz, 1 H)7.47-7.56 8.48 (NH): 1.10; 3.88; 2.53 5.10(OH): 3.57; 3.03 1.10 (Me):8.48; 3.88; 2.53 (m, 1 H) 7.59- 7.71 (m, 1 H) 8.35- 8.40 (m, 1 H) 8.48(s, 1 H) 8.79 (d, J = 2.20 Hz, 1 H) 9.16 (d, J = 2.20 Hz, 1 H) 85b

CIS-single stereoisomer b 1H N MR (500 MHz, DMSO-d6) δ ppm 0.94-1.07 (m,4 H) 1.12 (s, 3 H) 2.12 (tt, J = 8.38, 5.20 Hz, 1 H) 2.72 (br t, J =11.98 Hz, 1 H) 2.98 (br d, J = 13.69 Hz, 1 H) 3.47 (d, J = 11.25 Hz, 1H) 3.56 (br t, J = 11.74 Hz, 1 H) 3.75 (br dd, J = 11.37, 3.30 Hz, 1 H)3.81 (d, J = 11.25 Hz, 1 H) 5.38 (s, 1 H) 7.46- 7.56 (m, 2 H) 7.70 (t, J= 8.07 Hz, 1 H) 8.92 (NH): 5.38; 3.81; 3.56 5.38(OH): 8.92; 1.12 (Me):3.47; 2.72 8.35 (s, 1 H) 8.72 (d, J = 2.20 Hz, 1 H) 8.92 (s, 1 H) 9.12(d, J = 2.20 Hz, 1 H) 86b

TRANS-single stereoisomer b 1H NMR (500 MHz, DMSO-d6) δ ppm 0.95-1.07(m, 5 H) 1.10 (d, J = 2.57 Hz, 2 H) 2.12 (tt, J = 8.44, 5.14 Hz, 1 H)2.51- 2.56 (m, 1 H) 3.03 (td, J = 12.65, 4.28 Hz, 1 H) 3.54- 3.59 (m, 1H) 3.62 (t, J = 11.25 Hz, 1 H) 3.75 (br dd, J = 11.25, 2.45 Hz, 1 H)3.88 (d, J = 12.23 Hz, 1 H) 5.10 (s, 1 H) 7.46 (br d, J = 12.96 Hz, 1 H)7.47-7.56 8.48 (NH): 1.10; 3.88; 2.53 5.10(OH): 3.57; 3.03 1.10 (Me):8.48; 3.88; 2.53 (m, 1 H) 7.59- 7.71 (m, 1 H) 8.35- 8.40 (m, 1 H) 8.48(s, 1 H) 8.79 (d, J = 2.20 Hz, 1 H) 9.16 (d, J = 2.20 Hz, 1 H) 87a

TRANS-single stereoisomer a 1H NMR (400 MHz, DMSO-d6) δ ppm 2.41 (s, 3H) 2.47 (m, 1 H) 2.63- 2.74 (m, 1 H) 3.63-3.75 (m, 1 H) 3.76-3.87 (m, 2H) 3.94 (br d, J = 4.89 Hz, 1 H) 4.08 (d, J = 11.74 Hz, 1 H) 5.30 (d, J= 5.67 Hz, 1 H) 7.83 (d, J = 8.22 Hz, 1 H) 8.10 (dd, J = 8.31, 1.66 Hz,1 H) 8.38 (s, 1 H) 8.44 (s, 1 H) 8.82 8.44 (NH): 4.08; 3.94; 2.475.30(OH): 3.81, 2.68 3.94 (CH): 8.44; 2.47 (d, J = 1.96 Hz, 1 H) 8.85(d, J = 1.76 Hz, 1 H) 9.20 (dd, J = 1.96, 0.98 Hz, 1 H) 88a

TRANS-single stereoisomer a 1H NMR (500 MHz, DMSO-d6) δ ppm 0.87-0.99(m, 2 H) 1.00- 1.06 (m, 2 H) 2.10 (tt, J = 8.47, 5.10 Hz, 1 H) 2.46 (s,3 H) 2.51-2.53 (m, 1 H) 2.64-2.71 (m, 1 H) 3.64- 3.72 (m, 1 H) 3.77-3.88 (m, 3 H) 4.10 (d, J = 11.49 Hz, 1 H) 5.29 (br s, 1 H) 7.84 (d, J =8.07 Hz, 1 H) 8.08 (dd, J = 8.19, 2.08 Hz, 1 H) 8.65 (s, 1 H) 8.71 (d, —J = 2.20 Hz, 1 H) 8.83 (d, J = 2.20 Hz, 1 H) 9.04 (d, J = 1.71 Hz, 1 H)89b

TRANS-single stereoisomer b 1H NMR (500 MHz, DMSO-d6) δ ppm 0.92-1.07(m, 4 H) 2.12 (tt, J = 8.47, 5.10 Hz, 1 H) 2.44-2.48 (m, 1 H) 2.64-2.72(m, 1 H) 3.65- 3.73 (m, 1 H) 3.76- 3.86 (m, 2 H) 3.92 (br d, J = 5.87Hz, 1 H) 4.09 (d, J = 11.49 Hz, 1 H) 5.29 (d, J = 6.11 Hz, 1 H) 7.83 (d,J = 8.07 Hz, 1 H) 8.10 (dd, J = 8.19, 2.08 Hz, 1 H) 8.37 (s, 1 H) 8.43(s, 1 — H) 8.77 (d, J = 2.20 Hz, 1 H) 8.84 (d, J = 2.20 Hz, 1 H) 9.14(d, J = 2.20 Hz, 1 H)

1. A method for treating, ameliorating or preventing one or moreconditions comprising vascular dementia, age-associated memoryimpairment, cognitive impairment in bipolar disorder, cognitiveimpairment in schizoaffective disorder, cognitive impairment inHuntington's disease, cognitive impairment in autism spectrum disorders,cognitive impairment in Parkinson's disease, cognitive impairment afterstroke (post-stroke dementia), or cognitive impairment in vasculardementia in a subject comprising administering to the subject aneffective amount of a compound of formula (1)

wherein A is selected from the group consisting of

wherein above mentioned groups are substituted with one R⁵ and one R⁶;R¹ is selected from the group consisting of halogen, C₁₋₃-alkyl- andC₃₋₆-cycloalkyl- wherein the above mentioned C₁₋₃-alkyl-, andC₃₋₆-cycloalkyl-groups may optionally be substituted with 1 to 5substituents independently selected from the group consisting ofhalogen, NC— and HO—; R² is selected from the group consisting of aryland heteroaryl, wherein the above mentioned aryl and heteroaryl-groupsmay optionally be substituted with 1 to 5 substituents R⁴; R³ isselected from the group consisting of H— and C₁₋₃-alkyl-, wherein theabove mentioned C₁₋₃-alkyl-groups may optionally be substituted with 1to 7 substituents independently from each other selected from the groupconsisting of halogen; R⁴ is independently from each other selected fromthe group consisting of halogen, NC—, HO—, C₁₋₄-alkyl- and C₁₋₃-alkyl-O—wherein the above mentioned C₁₋₄-alkyl- and C₁₋₃-alkyl-O-groups mayoptionally be substituted with 1 to 5 substituents independentlyselected from the group consisting of HO— and F—; R⁵ is selected fromthe group consisting of H—, halogen, NC—, HO— and C₁₋₃-alkyl-, whereinthe above mentioned C₁₋₃-alkyl-group may optionally be substituted with1 to 5 substituents independently selected from the group consisting ofHO— and F— or R⁵ and R⁶ together form a group O═; R⁶ is selected fromthe group consisting of H—, halogen, NC—, HO— and C₁₋₃-alkyl-, whereinthe above mentioned C₁₋₃-alkyl-group may optionally be substituted with1 to 5 substituents independently selected from the group consisting ofHO— and F— or R⁵ and R⁶ together form a group O═; or a pharmaceuticallyacceptable salt thereof.
 2. The method of claim 1, wherein A is selectedfrom the group consisting of

wherein above mentioned groups are substituted with one R⁵ and one R⁶,or a pharmaceutically acceptable salt thereof.
 3. The method of claim 1,wherein A is selected from the group consisting of

or a pharmaceutically acceptable salt thereof.
 4. The method of claim 1,wherein A is selected from the group consisting of

or a pharmaceutically acceptable salt thereof.
 5. The method of claim 1,wherein R¹ is selected from the group consisting of F—, Cl—, C₁₋₃-alkyl-and C₃₋₆-cycloalkyl-, wherein the above mentioned C₁₋₃-alkyl- andC₃₋₆-cycloalkyl-groups may optionally be substituted with 1 to 3substituents independently selected from the group consisting of F—, ora pharmaceutically acceptable salt thereof.
 6. The method of claim 1,wherein R¹ is selected from the group consisting of F—, H₃C— andcyclopropyl-, or a pharmaceutically acceptable salt thereof.
 7. Themethod of claim 1, wherein R² is selected from the group consisting ofquinolinyl, phenyl and pyridynyl, wherein the above mentioned quinoline,phenyl and pyridyl-groups may optionally be substituted with 1 to 5substituents R⁴, or a pharmaceutically acceptable salt thereof.
 8. Themethod of claim 1, wherein R² is selected from the group consisting ofphenyl and pyridyl, wherein the above mentioned phenyl andpyridyl-groups may optionally be substituted with 1 to 2 substituentsR⁴, or a pharmaceutically acceptable salt thereof.
 9. The method ofclaim 1, wherein R³ is selected from the group consisting of H—, H₃C—,F₃C—, F₂HC—, FH₂C— and F₃C—, or a pharmaceutically acceptable saltthereof.
 10. The method of claim 1, wherein R⁴ is independently fromeach other selected from the group consisting of halogen, C₁₋₄-alkyl-and C₁₋₃-alkyl-O— wherein the above mentioned C₁₋₄-alkyl- andC₁₋₃-alkyl-O-groups may optionally be substituted with 1 to 5substituents independently selected from the group consisting of HO—,and F—, or a pharmaceutically acceptable salt thereof.
 11. The method ofclaim 1, wherein R⁵ is selected from the group consisting of H—, HO— andC₁₋₂-alkyl-, wherein the above mentioned C₁₋₂-alkyl-group may optionallybe substituted with 1 to 5 F—, or R⁵ and R⁶ together form a group O═, ora pharmaceutically acceptable salt thereof.
 12. The method of claim 1,wherein R⁶ is selected from the group consisting of H— and C₁₋₂-alkyl-,wherein the above mentioned C₁₋₂-alkyl-group may optionally besubstituted with 1 to 5 F—, or R⁵/R⁶ together form a group O═, or apharmaceutically acceptable salt thereof.
 13. The method of claim 1,wherein the compound of formula (1) is selected from the groupconsisting of:

or a pharmaceutically acceptable salt thereof.